U.S. patent application number 12/898313 was filed with the patent office on 2011-04-07 for treatment/cure of autoimmune disease.
This patent application is currently assigned to NUBIOME, INC.. Invention is credited to Richard Blankenbecler, Brian C. Lue, Fredrick C. Westall.
Application Number | 20110081320 12/898313 |
Document ID | / |
Family ID | 43823348 |
Filed Date | 2011-04-07 |
United States Patent
Application |
20110081320 |
Kind Code |
A1 |
Westall; Fredrick C. ; et
al. |
April 7, 2011 |
Treatment/Cure of Autoimmune Disease
Abstract
An embodiment entails administering an effective amount of a
product such as a protease or a source of a protease that destroys
or deactivates an immunogen, mimic or antigen specific to a
particular autoimmune disease before it encounters the immune
system of a patient.
Inventors: |
Westall; Fredrick C.;
(Temecula, CA) ; Blankenbecler; Richard; (Las
Vegas, NV) ; Lue; Brian C.; (Mountain View,
CA) |
Assignee: |
NUBIOME, INC.
Mountain View
CA
|
Family ID: |
43823348 |
Appl. No.: |
12/898313 |
Filed: |
October 5, 2010 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
|
61278333 |
Oct 6, 2009 |
|
|
|
Current U.S.
Class: |
424/93.4 ;
424/94.64; 424/94.67 |
Current CPC
Class: |
A61P 21/04 20180101;
A61K 38/4886 20130101; A61P 3/00 20180101; A61P 1/04 20180101; C12Y
304/24 20130101; A61K 38/482 20130101; A61P 19/02 20180101; A61P
25/00 20180101; A61P 29/00 20180101; C12Y 304/21026 20130101 |
Class at
Publication: |
424/93.4 ;
424/94.64; 424/94.67 |
International
Class: |
A61K 38/48 20060101
A61K038/48; A61P 25/00 20060101 A61P025/00; A61P 19/02 20060101
A61P019/02; A61P 21/04 20060101 A61P021/04; A61P 29/00 20060101
A61P029/00; A61P 1/04 20060101 A61P001/04; A61K 35/74 20060101
A61K035/74; A61P 3/00 20060101 A61P003/00 |
Claims
1. A method of treating Multiple Sclerosis, Rheumatoid Arthritis,
Anorexia Nervosa or Autistic Spectrum Disorder comprises:
administering an effective amount of a medicament comprised of
subtilisin to a mammal suffering from Multiple Sclerosis,
Rheumatoid Arthritis, Anorexia Nervosa or Autistic Spectrum
Disorder.
2. A method of treating Myasthenia Gravis, Uveoretinitis,
Ulcerative Colitis, Celiac Disease, Anorexia Nervosa or Autistic
Spectrum Disorder comprises: administering an effective amount of a
medicament comprised of oligopeptidase F (PepF) to a mammal
suffering from Myasthenia Gravis, Uveoretinitis, Ulcerative
Colitis, Celiac Disease, Anorexia Nervosa or Autistic Spectrum
Disorder.
3. A method of treating Myasthenia Gravis, Lupus Erythematosus,
Celiac Disease, Rheumatoid Arthritis or Autistic Spectrum Disorder
comprises: administering an effective amount of a medicament
comprised of endopeptidase O2 (PepO2) to a mammal suffering from
Myasthenia Gravis, Lupus Erythematosus, Celiac Disease, Rheumatoid
Arthritis or Autistic Spectrum Disorder.
4. A method of treating Uveoretinitis, Ulcerative Colitis, Celiac
Disease or Autistic Spectrum Disorder comprises: administering an
effective amount of a medicament comprised of endopeptidase O
(PepO) to a mammal suffering from Uveoretinitis, Ulcerative
Colitis, Celiac Disease or Autistic Spectrum Disorder.
5. A method of treating Celiac Disease or Autistic Spectrum
Disorder comprises: administering an effective amount of a
medicament comprised of Oenicoccus oeni to a mammal suffering from
Celiac Disease or Autistic Spectrum Disorder.
6. A method of treating Lupus Erythematosus or Rheumatoid Arthritis
comprises: administering an effective amount of a medicament
comprised of endopeptidase O from Bifidobacterium animalis subsp
lactis to a mammal suffering from Lupus Erythematosus or Rheumatoid
Arthritis.
7. The method of claim 5 wherein the medicament further comprises
endopeptidase O (PepO).
8. The method of claim 5 wherein the medicament further comprises
endopeptidase O2 (PepO2).
9. The method of claim 5 wherein the medicament further comprises
endopeptidase F (PepF).
Description
[0001] This patent application relates to U.S. Provisional
Application No. 61/278,333 filed Oct. 6, 2009 from which priority
is claimed under 35 USC .sctn.119(e), and which provisional
application is incorporated herein in its entirety.
TECHNICAL FIELD
[0002] One or more embodiments of the present invention provide
methods for treating or curing autoimmune diseases.
BACKGROUND
[0003] Chronic autoimmune diseases include Multiple Sclerosis,
Ulcerative Colitis, Lupus Erythematosus, Myasthenia Gravis,
Uveoretinitis, Arthritis, Autoimmune Diabetes, and Autoimmune
Neuritis, for example, Guillain Barre Syndrome. Many of these
diseases are characterized by unpredictable clinical exacerbations
and remissions.
[0004] These diseases may be initiated by microorganisms which
contain an amino acid sequence within one of its proteins that
mimics structurally a similar sequence within a protein in human
tissue (for example, see an article entitled "Sequence Homology
Between Certain Viral Proteins and Proteins Related to
Encephalomyelitis and Neuritis" by U. Jahnke, F. D. Fisher and E.
C. Alvord, Jr. in Science, 229 pp. 282-284 (1985) and an article
entitled "Amino Acid Homology Between the Encephalitogenic Site of
MBP and Virus: Mechanism for Autoimmunity" by R. S. Fujinami and M.
B. Oldstone in Science, 230 pp. 1043-1045 (1985)). When such a
microorganism invades an individual, it activates the immune system
against the mimic. The activated immune system then recognizes the
similar sequence within the protein in the human tissue, and
initiates an immune attack. This immune attack on its own tissue
results in the autoimmune disease.
[0005] A. Immunogens Producing Autoimmune Diseases: The identities
of immunogens, antigens, or mimics that trigger autoimmune diseases
are unknown. However, each human autoimmune disease has an animal
autoimmune disease counterpart. Further, a protein or proteins from
animal tissue that appear to be capable of activating these animal
autoimmune diseases have been identified. To test whether an agent
is an immunogen for the animal autoimmune disease, the suspected
agent is combined with an immunological adjuvant and injected into
a test animal. If clinical signs develop in the animal which are
similar to those for the human disease, the protein is considered
to be a potential immunogen responsible for the human disease. In
addition, regions, i.e., sequences, within these proteins have been
identified which themselves have been shown to trigger the
autoimmune response for the disease in animals. Further, the
contributions to induction of the autoimmune response of the
individual amino acids within these sequences have also been
determined. By knowing the contribution to disease induction of
each amino acid within an immunogen, one can identify potential
mimics.
[0006] B. Mimics: To identify a potential autoimmune mimic, one
first must have a defined immunogen. It is not sufficient that the
mimic have some or most of the same amino acids in its sequence.
Since each amino acid makes a different contribution towards the
autoimmune process, the mimic's sequence must match these
contributions. Table 1 in Appendix I lists a series of chemically
defined peptide immunogens which are able to initiate a variety of
experimental autoimmune diseases such as, for example, experimental
allergic encephalomyelitis (EAE)--a model for multiple sclerosis;
experimental allergic uveoretinitis (EAU)--a model for a variety
human retinitis maladies; experimental myasthenia gravis (EMG)--a
model for myasthenia gravis; and experimental systemic lupus
erythematosus (ELSE)--a model for lupus.
[0007] The immunogens responsible for experimental lupus contain a
high concentration of proline, e.g., PPPGMRPP. This, like the other
immunogens, should be particularly difficult to hydrolyze.
Furthermore it has been reported that in both the experimental
lupus and the corresponding human lupus, proteases involved in
proline peptide hydrolysis are depressed (for example, see
articles: "Abnormality of the post-proline cleaving enzyme activity
in mice with systemic lupus erythematosus-like syndrome" by T.
Aoyagi et al. in J. Appl. Biochem., 7 pp. 273-81 (1985); "Decreased
activities of serine proteinases in spleen of various murine models
of systemic lupus erythematodes" by T. Aoyagi, T. Wada, H. P.
Daskalov, F. Kojima, et al. in Biotechnol. Appl. Biochem., 9 pp.
362-7 (1987); "Dissociation between serine proteinases and proline
related enzymes in spleen of MRL mouse as a model of systemic lupus
erythematodes" by T. Aoyagi, T. Wada, H. P. Daskalov, F. Kojima, et
al. in Biochem. Int. 14 pp. 435-41 (1987); and "Activities of
Dipeptidyl Peptidase II and Dipeptidyl Peptidase IV in Mice and
Lupus Erythematosus-Like Syndrome and in Patients with Lupus
Erythematosus and Rheumatoid Arthritis" by M. Hagihara, M. Ohhashi
and T. Nagatsu in Clin. Chem., 33 pp 1463-1465 (1987).
[0008] Several other disease such as, ulcerative colitis, anorexia,
celiac disease, rheumatoid arthritis and even autism, are appear
also to have autoimmune aspects.
[0009] It is possible that no single etiological factor or agent is
responsible for the onset and perpetuation of ulcerative colitis.
However, some believe that ulcerative colitis is initiated by an
autoimmune response against some component of the intestinal
mucosa. An article by Das et al. (see an article entitled
"Autoimmunity to cytosketal protein tropomyosin. A clue to the
pathogenic mechanism for ulcerative colitis" by K. M. Das, A.
Dasgupta, A. Mandal, and X. Geng in J. Immunol., 150 pp. 2487-93
(1993)) has suggested that tropomyosin 3 isoform V, a 247 amino
acid protein, is such a component. An article by Mirza et al. (see
an article entitled "Autoimmunity Against Human Tropomyosin
Isoforms in Ulcerative Colitis" by Z. K. Mirza, S. Bhagyalakshmi,
J. J.-C. Lin, P. S. Amenta and K. M. Das in Inflamm. Bowel Dis., 12
pp. 1036-1043 (2006)) has shown that tropomyosin 3 isoform V is the
predominant isoform in the epithelium of colon and extra-intestinal
organs commonly involved in ulcerative colitis. An article by
Kesari et al. (see an article entitled "Externalization of
Tropomyosin Isoform 5 in Colon Epithelial Cells" by K. Y. Kesari,
N. Yoshizaki, X. Geng. J. J.-C. Lin and K. M. Das in Clin. Exp.
Immunol., 118 pp. 219-227 (1999)) has shown that tropomyosin 3
isoform V not only is located intracellularly within colon
epithelial cells, but more important, is normally released from
these cells into the colon. Released tropomyosin 3 isoform V can
interact with mimic activated gut immune cells, and the antibody
associated with tropomyosin blocks its activity in vitro. An
article by Das et al. (see an article entitled "Autoimmunity to
Cytosketal Protein Tropomyosin. A Clue to the Pathogenic Mechanism
for Ulcerative Colitis" by K. M. Das, A. Dasgupta, A. Mandal and X.
Geng in J. Immunol., 150 pp. 2487-93 (1993)) has suggested that
tropomyosin 3 isoform V plays an integral part in the pathology of
ulcerative colitis. An article by Geng et al. (see an article
entitled "Tropomyosin Isoforms in Intestinal Mucosa: Production of
Autoantibodies to Tropomyosin Isoforms in Ulcerative Colitis" by X.
Geng, L. Biancone, H. H. Dai, J. J. Lin, N. Yoshizaki, A. Dasgupta,
F. Pallone and K. M. Das in Gastrolenterol., 114 pp. 912-22 (1998))
has found antibodies directed against this protein in ulcerative
colitis in both the mucosa and sera. If its presence were used as a
diagnostic test to distinguish ulcerative colitis from Crohn's
disease, the test would have 100% specificity (see, for example, an
article entitled "Antibody to Tropomyosin Isoform 5 and Complement
Induce the Lysis of Coloncytes in Ulcerative Colitis" by E. C.
Ebert, X. Geng, M. Baipai, Z. Pan, E. Tater and K. M. Das in Am. J.
Gastroenterol., 104 pp. 2996-3003 (2009)). Autoantibodies against
human tropomyosin 3 isoform V in ulcerative colitis (see, for
example, an article entitled "Autoantibodies Against Human
Tropomyosin Isoform 5 in Ulcerative Colitis Destroys Colonic
Epithelial Cells Through Antibody and Compliment-Mediated Lysis" by
E. C. Ebert, X. Geng, J. Lin and K. M. Das in Cell. Immunol., 244
pp. 43-9 (2006)) destroy colonic epithelial cells through antibody
and complement-mediated lysis. An article by Vera et al. (see an
article entitled "Tropomodulin-Binding Site Mapped to Residues 7-14
at the N-Terminal Hepta Repeats of Tropomyosin Isoform 5" by C.
Vera, A. Sood, K. M. Gao, J. J. Lin and L. A. Sung in Arch.
Biochem. Biophys., 378 pp. 16-24 (2000)) has described a monoclonal
antibody which binds to the n-terminal region of tropomyosin,
blocking its ability to form with troponin as part of the membrane
complex. The binding site on tropomyosin that interacts with the
troponin is residues 7-14, IEAVRKKV. The binding site on the
tropomyosin that associates with the antibody is residues 4-10,
ITTIEAV. Mimics to this region could initiate an autoimmune
response.
[0010] There have been some molecular mimics suggested for MSH
alpha as a cause for anorexia. (see, for example, an article
entitled "Autoantibodies Against Appetite-Regulating Peptide
Hormones and Neuropeptides: Putative Modulation by Gut Microflora"
by S. O. Fetissov, S. M Hamze, M. Coeffer, et al., in Nutrition, 24
pp. 348-359 (2008)).
[0011] There are also some data involving proteolytic abnormalities
in human anorexia and bulimia. Two proteases, dipeptidyl peptidase
IV and prolyl endopeptidase, are abnormally low in anorexic and
bulimic patients (see, for example, articles: "Lowered Serum
Dipeptidyl Peptidase IV Activity in Patients with Anorexia and
Bulimia Nervosa" by D. van West, P. Monteleone, A. DiLieto, I de
Meister et al. in Eur. Arch. Psychiatry Clin. Neurosci., 250 pp.
86-92 (2000) and "Lower Serum Activity of Prolyl Endopeptidase in
Anorexia and Bulimia Nervosa" by M. Maes, P. Monteleone, R.
Bencivenga, F. Goossens, et al. in Psychoneuroendocrinolgy, 26 pp.
17-26 (2001)). Both enzymes are known to be involved in the
modification or inactivation of hormones operating in the
enteroinsular axis (see, for example, an article entitled "Eating
Disorders: A Role for Dipeptidyl Peptidase IV in Nutritional
Control" by M. Hildebrandt, M. Rose, H. Monnikes, W. Rutter, W.
Keller and B. F. Klapp in Nutrition, 17 pp. 451-4 (2001)). Both
proteases are involved in digestion of proline containing peptides
(see, for example, see article entitled "Isolation and
characterization of dipeptidyl peptidase IV from human placenta" by
G. Puschel, R. Mentlein and E. Heymann in Eur. J. Biochem., 126 pp.
359-65 (1982)).
[0012] Therapeutic Approaches for Autoimmune Diseases: Over the
last 50 years efforts have been undertaken to find treatments and
cures for chronic autoimmune diseases. Most of this work has
centered on finding general immunosuppressants.
[0013] Multiple Sclerosis (MS): MS is a chronic, slowly progressive
disease of the central nervous system characterized pathologically
by disseminated patches of demyelination in the brain and spinal
cord, and clinically by multiple symptoms and signs, for example,
paralysis, double vision, balance problems, and so forth, and by
remissions and exacerbations.
[0014] People have tried to treat/cure MS using a number of
different approaches. A first approach uses immunosuppressants.
Examples include: ACTH (see, for example: an article entitled
"Adrenal steroid therapy in neurological disease" by A. A. Eisen
and J. W. Norris in Canad. Med. Ass. J., 100 pp. 27-30 (1969));
cyclophosphamide (see, for example, an article entitled "Intensive
immunosuppression with cyclophosphamide in multiple sclerosis.
Follow up of 110 patients for 2-6 years" by R. E. Gonsette, L.
Demonty and P. Delmotte in J. Neurol., 214 pp 173-81 (1977));
prednisone (see, for example, an article entitled: "Prolonged
effects of large-dose methylprednisolone infusion in multiple
sclerosis" by J. L. Trotter and W. F. Garvey in Neurol., 30 pp.
702-8 (1980)); azothioprine (see, for example, an article entitled
"Long-term treatment of multiple sclerosis with azathioprine" by W.
R. Swinburn and L. A Liversedge in J. Neurol. Neurolsurg. Psychia.,
36 pp. 124-126 (1973)); beta interferon (see, for example, an
article entitled "Double blind study of intrathecal beta-interferon
in multiple sclerosis: clinical and laboratory results" by C.
Milanese, A. Salmaggi, L. La Mantia, A. Campi, M. Eoli, et al. in
J. Neurol. Neurosurg. Psychia., 53 pp. 554-557 (1990)); interferon
(see, for example, an article entitled "Multicentre double-blind
study of effect of intrathecally administered natural human
fibroblast interferon on exacerbations of multiple sclerosis" by L.
Jacobs, A. M. Salazar, R. Herndon, P. A. Reese, et al. in Lancet, 2
pp. 1411-3 (1986)); and mitoxantrone (see, for example, an article
entitled "Treatment of multiple sclerosis with mitoxantrone" by E.
Mauch, H. H. Kornhuber, H. Krapf, U. Fetzer and H. Laufen et al. in
Eur. Arch. Psychia Clin. Neurosci., 244 pp. 96-102 (1992)). Other
procedures have been used to generally depress the immune system of
multiple sclerosis patients. These include: plasmapheresis (see,
for example, an article entitled "Plasmapheresis in multiple
sclerosis: preliminary study" by H. L. Weiner and D. M. Dawson in
Neurol., 30 pp. 1029-33 (1980)); use of antithymocyte globulin
(see, for example, an article entitled "Multiple sclerosis treated
with antithymocyte globulin--a five year follow-up" by L. K.
Kastrukoff, D. R. McLean and T. A. McPherson in Can. J. Neurol.
Sci., 5 pp. 175-8 (1978)); use of various antibody concoctions
(see, for example, articles: "Immunogenic responses of progressive
multiple sclerosis patients treated with an anti-T-Cell monoclonal
antibody, anti-T12" by D. A. Hafler, R. J. Fallis, D. M Dawson, S.
F. Scholssman et al in Neurol., 36 pp. 777-84 (1986); "An open
trial of OKT3 in patients with multiple sclerosis" by B. G.
Weinshenker, B. Bass, S. Karlik, G. C. Ebers and G. P. Rice in
Neurol., 41 pp. 1047-52 (1991); and "Treating multiple sclerosis
with monoclonal antibodies: a 2010 update" by M. Buttmann in Expert
Rev. Neurother., 10 pp. 791-809 (2010)); use of a transfer factor
(see, for example, an article entitled "A double blind trial of
transfer factor vs. placebo in multiple sclerosis patients" by R.
C. Collins, L. R. Espinoza, C. R. Plank, G. C. Ebers et al. in
Clin. Exp. Immunol., 33 pp. 1-11 (1978)); and stem cell
transplantation (see, for example, an article entitled "Stem cell
transplantation in multiple sclerosis: current status and future
prospects" by G. Martino, R. J Franklin, A. B. van Evercooren, and
D. A. Kerr in Nat. Rev. Neurol., 6 pp. 247-55 (2010)). However,
these human autoimmune diseases are chronic diseases. In many cases
they have unpredictable relapsing and remitting episodes.
Therefore, predicting when an immunosuppressive should be
administered is questionable. In any event, the patient would be
"exposed" to immunosuppressants throughout life. Therefore, the
patient will have an increased susceptibility to any and all other
infective agents.
[0015] Another approach is to eliminate the autoimmune response
against the agent responsible, i.e., the mimic or immunogen, for
the disease with a vaccine. This involves multiple injections of
high concentrations of the immunogen, intending to deactivate the
immune system against the immunogen--this is called antigen
suppression. This approach has been used clinically in humans or in
animals. An advantage of this approach is that the immune system is
not totally immunosuppressed, but it is only suppressed against a
particular autoimmune immunogen. A disadvantage is that one needs
to identify precisely the immunogen. In most cases this is quite
difficult. For example, four separate proteins have been identified
as potential MS immunogens. Within these four proteins, several
regions have been suggested as potential MS immunogens. Thus, there
are potentially 20 different MS sites. This makes it difficult for
this approach to succeed.
[0016] One of the proteins implicated in Multiple Sclerosis is the
human myelin basic protein. Dr. Jonas Salk (see, for example, an
article entitled "Myelin Basic Protein Treatment of MS" by J. Salk,
F. Westall, J. Romine and W. Wiederholt in Neurol., 29 pp. 573
(1979)) attempted to cure multiple sclerosis with multiple
injections of high concentrations of porcine myelin basic protein
(MBP) and failed. When comparing the sequence of porcine MBP to the
human MBP, one finds a major sequence change in one of the
immunogenic regions. A similar therapy involves injection of a
copolymer of four amino acids which represent four amino acids
involved in the induction of allergic encephalomyelitis.
Unfortunately, the co-polymer was modeled after an immunogen which,
from the Salk experiment described above, was shown not to be not
the MS immunogen. In fact, the co-polymer has been used with little
success for over 20 years (see, for example, an article entitled
"Glatiramer acetate and the glatiramoid class of immunomodulator
drugs in multiple sclerosis: an update" by K. P. Joohnson in Expert
Opin. Drug Metab. Toxicol., 6 pp 643-60 (2010)).
[0017] In other approaches, therapies have also been developed to
alleviate symptoms rather than to eradicate the causes of the
symptoms. These include: the use of brolitene (see, for example, an
article entitled "A Trial of Brolitene in the Treatment of
Spasticity" by G. D. Perkin and M. J. Aminoff in Br. J. Clin.
Pharmac., 3 pp. 879-882 (1976)); the use of bactofen (see, for
example, an article entitled "A double blind trial with bactofen
and diazepam in spascity due to multiple sclerosis" by A. From and
A. Heltberg in Acta Neurol. Scand., 51 pp. 158-66 (1975)); the use
of carbamazeprine (see, for example, an article entitled "Treatment
of paroxysmal disorders in multiple sclerosis with carbamazepine"
by M. L. E. Espir and P. Millac in J. Neurol. Neurosurg. Psychiat.,
33 pp. 528-531 (1970)), the use of glutethimide (see, for example,
an article entitled "Glutethimide treatment of disabling action
tremor in patients with multiple sclerosis and traumatic brain
injury" by M Aisen, M Holzeer, M Rosen and M Dietz in Arch.
Neurol., 48 pp. 513-5 (1991)); the use of 9-tetrahydrocannabinol
(see, for example, an article entitled "Treatment of human spascity
with delta 9-tetrahydrocannabinol" by D. J. Petro and C.
Ellenberger, Jr. in J. Clin. Pharmacol., 21 pp. 413S-416S (1981));
the use of 4-aminopyridine (see, for example, an article entitled
"4-aminopyridine in multiple sclerosis: prolonged administration"
by D. Stefoski, F. A. Davis, W. E. Fitzsimmons, S. S. Luskin et al.
in Neurol., 441 pp. 1344-8 (1991)); and the use of dantrolene (see,
for example, an article entitled "The effects of dantrolene sodium
in relation to blood levels in spastic patients after prolonged
administration" by W. J. Meyler, H. Bakker, J. J. Kok, S. Agoston
and H. Wesseling in J. Neurol. Neurolsurg. Psychia., 44 pp. 334-339
(1981)).
[0018] Further approaches include: use of hyperbaric oxygen (see,
for example, an article entitled "Hyperbaric oxygen in multiple
sclerosis: a double blind trial" by C. M. Wiles, C. R. A. Clarke,
H. P. Irwin, E. F. Edgar and A. V. Swan in Brit. Med. J., 292 pp.
367-371 (1986)); linoelate supplementation (see, for example, an
article entitled "Double blind trial of linoleate supplementation
of the diet of multiple sclerosis" by J. H. D. Millar, K. J.
Zilkha, M. J. S. Langman, H. P. Wright, et al. in Brit. Med. J., 1
pp. 765-768 (1973)); and the use of diets (see, for example,
articles: "Letter: Gluten free diet as treatment for multiple
sclerosis" F. H. Hafner in Postgrad. Med., 59 p. 20 (1976); "Review
of MS patient survival on a Swank low saturated fat diet" by R. L.
Swank and J. Goodwin in Nutrition, 19 pp. 161-2 (2003); and "A
double blind controlled trial of long chain n-3 polyunsaturated
fatty acids in the treatment of multiple sclerosis" by D. Bates, N.
E. F. Cartlidge, J. M. French, M. J. Jackson, et al. in J. Neurol.
Neurolsurg. Psycia., 52 pp. 18-22 (1989)). These approaches have
provided little long term success.
[0019] Lastly, in another approach, O. A. Mlalovyts'ka reports a
therapy entailing the use of various enzymes (see the abstract of
an article (in Ukrainian) entitled "Effect of phlogenzym in
long-term treatment of patients with multiple sclerosis" by O. A.
Mlalovyts'ka in Lik. Sprava., pp. 109-13 (April-June, 2003)). The
abstract indicates that Phlogenzym, a product comprised of the
enzymes bromelain and trypsin and the anti-oxidant rutin were
administered orally to 74 patients with progressing MS. The
abstract indicates that over a one to three year period, their
symptoms gradually improved and the rate of progression slowed.
[0020] Myasthenia Gravis: Myasthenia gravis is characterized by
progressive fatigability of muscles due to impairment of conduction
at the myoneural junction. Patients complain of double vision,
difficulty swallowing, chewing and talking. The course is variable
and there may be prolonged remissions.
[0021] People have tried to treat/cure Myasthenia Gravis using
immunosuppressants such as corticosteroids that reduce inflammatory
signaling molecules and adhesion molecules and reduce the transport
of inflammatory cells. For example, prednisone has been used (see,
for example, an article entitled "Treatment of autoimmune
myasthenia gravis" by D. Richman and M. Agius in Neurology, 61 pp.
1652-1661 (2003)). In general, this approach induces remission in
many patients, but long-term usage has serious side-effects, and
does not cure the disease.
[0022] Uveoretinitis: Uveoretinitis encompasses a collection of
human sight-threatening inflammatory eye diseases of autoimmune
etiology. Experimental uveoretinitis is the model for the
experimental model for these diseases.
[0023] The use of immunosuppressants is a standard procedure for
treating uveoretinitis. Such procedures include: use of adalimumab
(see, for example, an article entitled "Adalimumab therapy for
refractory uveitis: a pilot study" by M. S. Diaz-Liopis, S.
Garia-Delpech, D. Salom, P. Udaondo, et al. J. Ocul. Pharmacol.
Ther., 24 pp. 351-61 (2008)); use of dexamethasone (see, for
example, an article entitled "Dexamethasone sustained drug delivery
implant for the treatment of severe uveitis" by G. J. Jaffe, P. A.
Pearson and P. Ashton in Retina 20 pp. 402-3 (2000)); use of
guanethidine (see, for example, an article entitled "Guanethidine
in the therapy of glaucoma" by J. A. Castren, S. Pohjola, P.
Pakarinen and K. Karjalainen in Ophthalmologica, 155 pp. 194-204
1968); use of infliximab (see, for example, an article entitled
"Treatment of refractory posterior uveitis with infliximab: a 7
year follow study" by R. Lopez-Gonzalez, E. Loza, J. A. Jover, J.
M. Benitez del Catillo, et al. in Scand. J. Rheumatol., 38 pp.
58-62 (2009)); and use of cyclosporine (see, for example, an
article entitled "Cyclosporine therapy for uveitis: long-term
follow up" by R. B. Nussenblatt, A. G. Palestine and C. C. Chan in
J. Ocul. Pharmacol., 1(4) pp. 369-82 (1985)). Oral administration
of retinal antigens has also been used to attempt specific antigen
suppression (see, for example, an article entitled "Treatment of
uveitis by oral administration of retinal antigens: results of a
phase I/II randomized masked trial." by R. B. Nussenblatt, I. Gery,
H. L. Weiner, F. L. Ferris, J. Shiloach, et al. in Am. J.
Ophthalmol., 123 pp. 583-92 (1997)). The use of interferon alpha 2a
(see, for example, an article entitled "Interferon alpha-2a: a new
treatment option for long lasting refractory cystoid macular edema
in uveitis? A pilot study" by C. M. Deuter, I. Koetter, I.
Guenaydin, N. Stuebiger and M. Zierhut in Retina 26 pp. 786-91
(2006)) controls various secondary problems associated with the
disease, including macular edema. Another therapy entails
cryotherapy (see, for example, an article entitled "Treatment of
peripheral uveoretintitis by cryotherapy" by T. M. Aaberg, T. J.
Cesarz and R. R. Flickinger in Am. J. Ophthalmol., 75 pp. 685-8
(1973)). In addition, the use of certain immunomodulator drugs has
been proposed (see, for example, an article entitled "The potential
of newer immunomodulating drugs in the treatment of uveitis: a
review" by J. Salzmann, and S. Lightman in BioDrugs, 13 pp. 397-408
(2000)).
[0024] Lupus Erythematosus: Lupus erythematosus is a collection of
chronic systemic autoimmune diseases--four main types of lupus
exist. Of these, systemic lupus erythematosus is the most serious.
Symptoms vary from person to person. Almost all people with
systemic lupus erythematosus have joint pain and swelling--some
develop arthritis. Other major symptoms include: fatigue, fever,
mouth sores, hair loss, sensitivity to light, skin rashes and chest
pains.
[0025] People have tried to treat/cure Lupus erythematosus using a
number of different approaches. A first approach entails
supplementation of beneficial bacteria, known as probiotics, to
compete with the effects of suspected harmful bacteria, such as
Lactobacillus bulgaricus delbruekii and Streptococcus thermophilus
(see U.S. Pat. No. 6,696,057 entitled "Composition and method for
treatment of gastrointestinal disorders and hyperlipidemia"). Use
of this approach has been intended to reduce symptoms associated
with the disease, but there have been no data published to support
this.
[0026] Another approach entails delivering human recombinant Dnase
1 intravenously or subcutaneously. Human recombinant Dnase 1 is an
enzyme that breaks down excess DNA fragments found in serum (see,
for example, a book entitled "Dubios' lupus erythematosus," by D.
Wallace, B. Hahn and E. Dubois at p. 1354, Lippincott Williams
& Wilkins (2007)). In general, this approach has not been
successful at treating or curing the disease.
[0027] Ulcerative Colitis (UC): UC is a chronic, nonspecific,
inflammatory and ulcerative disease of the colon which is
characterized by the passage of bloody and purulent mucus, either
alone or accompanying formed or watery stools. The cause of UC is
unknown.
[0028] People have tried to treat/cure UC using a number of
different approaches. A first approach entails giving patients
antibiotics to kill microbes that may be causing or exacerbating
symptoms. For example, the following antibiotics have been used:
clarithromycin, ethambutol, metronidazole, and rifabutin (see, for
example, an article entitled "Treatment of Ulcerative Colitis Using
Fecal Bacteriotherapy" by T. Borody, E. Warren, S. Leis, R. Surace
and O. Ashman in J. Clin. Gastroenterol., 37(1) pp. 42-47 (2003)).
Although antibiotics can help treat the symptoms of ulcerative
colitis, in general, this approach has not been successful at
curing the disease. A second approach entails treating patients
with various immunosuppressants such as azothioprine, prednisone,
prednisolone sodium phosphate enema, mercaptopurine, cyclosporine,
and clofazimine (see, for example, an article entitled "Treatment
of Ulcerative Colitis Using Fecal Bacteriotherapy" by T. Borody, E.
Warren, S. Leis, R. Surace and O. Ashman in J. Clin.
Gastroenterol., 37(1) pp. 42-47 (2003)). Although these drugs
reduce symptoms, they have significant side effects for long-term
use. Another approach entails treating patients with various
anti-Tumor Necrosis Factor antibodies such as infliximab (see, for
example, an article entitled "Review article: Infliximab therapy
for inflammatory bowel disease--seven years on" by P. Rutgeerts, G.
Van Assche, and S. Vermeire in Aliment Pharmacol. Ther., 23(4) pp.
451-63 (Feb. 15, 2006)). However, not all people respond to
anti-Tumor Necrosis Factor antibodies, some have allergic
reactions, and their use increases the risk of infection. Another
approach suppresses the immune system by suppressing inflammatory
signaling molecules, thereby slowing down production of immune
cells, using anti-tumor necrosis factor drugs to block TNF
activity. For example, the following have used: clarithromycin,
ethambutol, metronidazole, mercaptopurine, and rifabutin (see, for
example, an article entitled "Treatment of Ulcerative Colitis Using
Fecal Bacteriotherapy" by T. Borody, E. Warren, S. Leis, R. Surace
and O. Ashman in J. Clin. Gastroenterol., 37(1) pp. 42-47 (2003)).
In general, this approach can treat with the increased risk of
infection and other side-effects, but not cure the disease.
[0029] Another approach entails treating with synthesized
anti-oxidant chemicals to block free-radicals from damaging
gastrointestinal tissues. These chemicals, called aminosalicylates,
include mesalamine, olsalazine and salazopyrin (see, for example,
an article entitled "Treatment of Ulcerative Colitis Using Fecal
Bacteriotherapy" by T. Borody, E. Warren, S. Leis, R. Surace and O,
Ashman in J. Clin. Gastroenterol., 37(1) pp. 42-47 (2003)). In
general, this approach treats symptoms of some patients but does
not, in general, cure the disease. Another approach entails
treating with various anti-motility drugs such as loperamide
hydrochloride and codeine phosphate to reduce diarrhea (see, for
example, an article entitled "Treatment of Ulcerative Colitis Using
Fecal Bacteriotherapy" by T. Borody, E. Warren, S. Leis, R. Surace,
and O. Ashman in J Clin Gastroenterol 37(1) pp. 42-47 (2003)). In
general, this reduces the frequency of stools, but it has a risk of
inducing toxic megacolon associated with the disease. Another
approach entails treating with various enzymes extracted from
plants, such as bromelain (see, for example, an article entitled
"Use of Bromelain for Mild Ulcerative Colitis" by S. Kane and M.
Goldberg in Annals of Internal Medicine, vol. 132, no. 8 p. 680
(Apr. 18, 2000)). Although the article states "Orally administered
bromelain was anecdotally reported to induce clinical and
endoscopic remission of ulcerative colitis in two patients whose
disease was refractory to multi-agent conventional medical
therapy," in general, this approach has not been successful at
treating or curing the disease in significant numbers of
patients.
[0030] Another approach entails treating with short chain fatty
acids such as butyric acid in enemas to rapidly nourish colonic
tissues (see an article entitled "Local Short-Chain Fatty Acids
Supplementation without Beneficial Effect on Inflammation in
Excluded Rectum" by J. Schauber, T. Bark, E. Jaramillo, M. Katouli,
B. Sandstedt and T. Svenberg in Scand. J. Gastroenterol., (2)
(2000)). In general, this approach has not been successful at
treating or curing the disease. Another approach entails treating
with bismuth compounds such as oral bismuth subsalicylate to
chemically remove toxic hydrogen sulfide from the intestine (see,
for example, an article entitled "Production and elimination of
sulfur-containing gases in the rat colon" by F. Suarez, J. Fume, J.
Springfield and M. Levitt in Am. J. Physiol., 274(4 Pt 1): pp.
G727-33 (April 1998)). In general, this approach has not been
successful at treating or curing the disease. Another approach
entails treating with anti-inflammatory compounds from plants such
as aloe vera gel (see, for example, an article entitled
"Randomized, double-blind, placebo controlled trial of aloe vera
gel for active ulcerative colitis" by L. Langmead, R. Feakins, S.
Goldthorpe, H. Holt, E. Tsironi, A. De Silva, D. Jewell and D.
Rampton in Aliment Pharmacol. Ther., 19 pp. 739-747 (2004)).
Although this approach has been able to reduce symptoms, the
results have not shown to be successful at treating or curing the
disease in large numbers of patients over long periods of time.
[0031] Another approach entails treating by supplementing the diet
with substances to promote the growth of beneficial bacteria,
called prebiotics, such as germinated barley foodstuff (see, for
example, an article entitled "The Dietary Combination of Germinated
Barley Foodstuff plus Clostridium butyricum Suppresses the Dextran
Sulfate Sodium-Induced Experimental Colitis in Rats" by Y. Araki,
Y. Fujiyama, A. Ando, S. Koyama, O. Kanauchi and T. Bamba in Scand.
J. Gastroenterol., 35 (10) pp. 1060-7 (October 2000)). In general,
this approach has been successful at helping patients go into, and
maintain, remission longer than a placebo. However, it has not been
shown to stop periodic flare-ups associated with the disease.
Another approach entails treating by infecting the patient with
intestinal worms to change the immune behavior from Th1 to Th2
which reduces inflammation in the colon (see, for example, an
article entitled "What is the origin of ulcerative colitis? Still
more questions than answers" by M. Lukas, M. Bortilik and Z.
Maratka in Postgrad. Med. J., 82 pp. 620-625 (2006)). In general,
this approach has helped treat patients' symptoms, but does not
cure the disease. Another approach entails treating by
supplementing the diet with colonically delivered
phosphatidylcholine which is believed to make the colon hydrophobic
and less vulnerable to toxins (see, for example, an article
entitled "Retarded release phosphatidylcholine benefits patients
with chronic active ulcerative colitis" by W. Stremmel, U. Merle,
A. Zahn, F. Autschbach, U. Hinz and R. Ehehalt in Gut, 54 pp.
966-971 (2005)). Although this approach helped many patients
achieve remission, a significant number complained of side-effects,
bloating and nausea in a later study.
[0032] Another approach entails treating by supplementation of
beneficial bacteria, known as probiotics, to compete with the
effects of suspected harmful bacteria, such as Escherichia coli
Nissle 1917 (see, for example, an article entitled "Bacteria as the
cause of ulcerative colitis" by M. Campieri and P. Gionchetti in
Gut, 48(1) pp. 132-5 (January 2001)). Although this approach has
been shown to reduce symptoms and lengthen the time between
flare-ups of the disease, it could not prevent periodic flares that
occur about once a year in large numbers of people. Another
approach entails treating by replacing bacteria in the
gastrointestinal tract with a human fecal transplant (see, for
example, an article entitled "Treatment of Ulcerative Colitis Using
Fecal Bacteriotherapy" by T. Borody, E. Warren, Sh. Leis, R. Surace
and O. Ashman in J. Clin. Gastroenterol., 37(1) pp. 42-47 (2003)).
In general, although this approach helped mitigate patients'
symptoms, it has not been successful at curing the disease in all
patients.
[0033] Celiac Disease (CD): CD is a fairly common genetically
determined inflammatory disease of the intestine, and is induced by
ingestion of gluten and related proteins from barley, oats and rye.
Symptoms are age dependent. Children suffer from diarrhea, fatigue
and weight loss; in older individuals symptoms are more diffuse.
Both immune responses to gluten and autoimmune triggers are thought
to be involved in the disease. In active CD, the small intestinal
mucosa is affected by both innate and adaptive immune processes.
Inflammatory lesions occur in the upper small intestinal mucosa
characterized by increased numbers of intraepithelial lymphocytes
together with small intestinal villous atrophy and crypt
hyperplasia.
[0034] People have tried to treat/cure CD with various enzymes
extracted from plants and microbes such as a cysteine endoprotease
from germinating barley and prolyl endopeptidase from Sphingomonas
capsulata (see, for example, an article entitled "The effects of
ALV003 pre-digestion of gluten on immune response and symptoms in
celiac disease in vivo" by J. Tye-Din, R. Anderson, R. French, G.
Brown, P. Hodsman, M. Siegel, W. Botwick and R. Shreeniwas in
Clinical Immunology, vol. 134 pp. 289-295 (2010)). Although the
article states that the enzymes were successful at destroying
targeted gluten peptides and prevented an immune response to
gliadin or 33 mer, the patients' symptoms did not improve and even
worsened during the clinical trial. This indicates that, in
general, the approach has not been successful at treating or curing
the disease in significant numbers of patients.
[0035] Anorexia nervosa: Anorexia nervosa is an eating disorder
characterized by refusal to maintain a healthy body weight and an
obsessive fear of gaining weight. People have tried to treat/cure
anorexia nervosa with antidepressants such as olanzapine (see, for
example, an article entitled "Review of Medication Use for Children
and Adolescents with Eating Disorders" by J. Couturier and J. Lock
in J. Can. Acad. Child. Adolesc. Psychiatry, 16:4 (November 2007)).
In general, this approach can treat symptoms but does not cure the
disease.
[0036] Rheumatoid Arthritis: Rheumatoid arthritis is an autoimmune
disease characterized by chronic inflammation of the synovial
membrane, resulting in destruction of cartilage and bone in
affected joints. People have tried to treat/cure rheumatoid
arthritis using a number of different approaches. A first approach
entails giving patients antibiotics to kill microbes that may be
causing or exacerbating symptoms. For example, the antibiotic
minocyclin has been used (see, for example, an article entitled
"Use of aminocycline in rheumatoid arthritis: a district general
hospital experience" by E. Suresh, I. M. Morris and P. C. Mattingly
in Ann. Rheum. Dis., 63 pp. 1354-1355 (2004)). Although the
treatment helped some patients reduce their symptoms, many had
substantial side effects and had to stop treatment. In general,
this approach has not been successful at curing the disease. A
second approach entails giving patients various immunosuppressants
such as prednisone, methotrexate, infliximab, adalimumab and
entanercept. (see, for example, an article entitled "Serious
infections with antirheumatic therapy: are biologicals worse?" by
K. L Winthrop in Ann. Rheum. Dis., 65(Suppl III) pp. iii54-iii57
(2006)). Although these chemicals are sometimes effective at
reducing symptoms, they have significant side-effects for long term
use. In general, this approach has been successful at treating,
with risk of infection, but not at curing the disease. Another
approach entails the use of various enzymes (see, for example, an
article entitled "Basic studies on enzyme therapy of immune complex
diseases" by C. Steffen and J. Menzel in Wein. Klin. Wochenschr.,
97(8) pp. 376-85 (Apr. 12, 1985)). The abstract indicates that
several unidentified enzymes were administered orally to rabbits
whose arthritis was induced experimentally with antigens and that
the enzymes helped reduce symptoms. Because this was an animal
study, there is no data to indicate if the approach can be
successful at treating or curing the disease in significant numbers
of patients.
[0037] Autoimmune Autistic Spectrum Disorder: There are many causes
of autistic spectrum disorders and research has shown that a
history of autoimmunity, especially Celiac disease, ulcerative
colitis, and rheumatoid arthritis in the mother are risk factors
for the child to become diagnosed with an autistic spectrum
disorder. The causes and treatments for autistic spectrum disorder
people are highly varied. Some are sensitive to gluten and casein
and respond to dietary elimination of gluten and casein; some
respond to antibiotics, such as oral vancomyocin, but their
symptoms return soon after stopping consumption of the antibiotic;
some respond to dietary supplementation with probiotics as well as
enzymes. One doctor announced that he had observed favorable
responses with an enzyme sold under the trade name Nattokinase.
Lactobacilli, Bifidobacterium, Lactococcus, and Saccharomyces
boulardii are probiotics that are often recommended by doctors to
autistic children who have gastrointestinal issues such as diarrhea
or constipation.
SUMMARY
[0038] One or more embodiments of the present invention comprise
destroying or deactivating an immunogen, mimic and/or antigen for a
disease before it encounters the immune system. In particular, an
embodiment of one method comprises administering a source of a
protease that destroys or deactivates an immunogen, mimic and/or
antigen specific to a particular autoimmune disease before it
encounters the immune system.
DETAILED DESCRIPTION
[0039] One or more embodiments of the invention are methods for
delivering or administering a medicament comprised of protolytic
enzymes, as chemical compounds, or for delivering or administering
a medicament comprised of probiotic microorganisms that produce
such protolytic enzymes, which protolytic enzymes eradicate or
degrade autoimmune mimics to prevent the interaction of the
autoimmune mimics with a host's (otherwise referred to herein as a
patient) patient immune system--such hosts being mammals such as,
for example and without limitation, humans, dogs, cats and so
forth. As used herein, autoimmune mimics are structurally
comparable to autoimmune immunogens found in tissues of hosts
afflicted with an autoimmune disease. In accordance with one or
more such embodiments, the proteolytic enzymes are delivered to the
gastrointesintal tract by probiotic microorganisms, by a drug
delivery system, or by a combination of enzymic chemistries and
probiotic microorganisms. As used herein, the term host also refers
to a patient.
[0040] In accordance with one or more embodiments, the proteolytic
enzymes include selected proteases which can be delivered: (a) as
pure chemicals; or (b) as pure chemicals in combination with live
and/or dead microorganisms or portions of such microorganisms. In
accordance with one or more such embodiments, the selected
protease(s) will reduce the concentrations of peptides in a host's
gut which are composed of more than six (6) amino acids. As such,
the selected protease(s) will reduce the concentration of potential
autoimmune mimics already present in a host's inflicted gut, and
prevent new autoimmune mimics from being produced. Advantageously,
as a result, the probiotic microorganisms and/or proteases will
eliminate potential autoimmune mimics to provide a procedure for
treating/curing chronic autoimmune diseases.
[0041] In accordance with one or more embodiments, a medicament
comprised of probiotic microorganisms or proteases or a combination
of probiotic microorganisms and proteases may be delivered or
administered orally, by suppository, or by injection into a
patient's gut (for example, and without limitation, enema,
endoscope, colonoscope, robotically actuated capsule, and so
forth). In accordance with one or more such embodiments, treatment
would range from about weekly to about daily, and be ongoing until
symptoms of the disease have disappeared.
[0042] Multiple Sclerosis (MS): In accordance with one or more
embodiments of the present invention, a medicament comprised of an
effective dose or amount of subtilisin is administered to a patient
to treat/cure MS. An effective dose or amount of subtilisin is a
dose or amount of the protease that is effective in destroying or
deactivating mimics, immunogens and/or antigens that cause or
exacerbate MS. The effective amount of subtilisin administered will
depend upon the severity of the disease process, but will range
between about 2,000 fibrinolytic units/day and about 10,000
fibrinolytic units/day. For example, the subtilisin will be
administered in one or more, preferably three, doses daily of
subtilisin. It is believed that hydrolytic processes of the
subtilisin will act in two areas: the gut and the blood. In the
gut, the subtilisin will destroy or deactivate potential MS mimics
before they can reach a sufficient level for immune activation,
and, in the blood the subtilisin will destroy or deactivate any MS
immunogens and antigens that are released from central nervous
system myelin which could interact with the immune system and
prolong a clinical exacerbation.
[0043] In accordance with one or more such embodiments of the
present invention, a medicament comprised of an effective amount of
a non-pathogenic microorganism and/or its spore (that are capable
of providing subtilisin), is administered to a patient to
treat/cure MS. In accordance with one or more such embodiments,
suitable microorganisms and spores include, for example, but not
limited to, Bacillus subtilis, Bacillus licheniformis, and Bacillus
lentus and their various strains. An effective amount of the
microorganism and/or its spores is an amount that is effective to
cause destruction or deactivation of mimics, immunogens and/or
antigens that cause or exacerbate MS. In accordance with one or
more such embodiments, an effective amount is from about 100
thousand CFU to about 600 billion CFU per dose (CFU designates
colony forming units), where the dose is administered about one or
more times per week, or as often as about one to about three times
daily.
[0044] In accordance with one or more embodiments of the present
invention, a medicament comprised of an effective amount of parts
of, or entire broken up, microorganisms and/or their spores capable
of providing subtilisin, is administered to a patient to treat/cure
MS. Parts of the microorganisms and/or spores can be separated and
selected, using any one of a number of methods that are well known
to those of ordinary skill in the art, for their bioactive
properties to help ensure and improve the rate of the destruction
or deactivation of mimics, immunogens or immunogens that cause or
exacerbate MS and/or to improve the effectiveness of enzymes in the
gastrointestinal or respiratory tract in destroying or deactivating
such mimics, immunogens and/or antigens. As used herein, the term
gastrointestinal tract includes the oral cavity. In accordance with
one or more embodiments, the parts of the microorganisms or spores
selected can be, but are not limited to, adjuvants such as muramyl
dipeptide. An adjuvant's content can be increased or decreased
using any one of a number of methods that are well known to those
of ordinary skill in the art to saturate or starve the
gastrointestinal tract or respiratory tract of the adjuvant to
prevent an optimum ratio of adjuvant to mimic from being close
enough to one that initiates and/or maintains the autoimmune
reaction. In accordance with one or more embodiments, suitable
microorganisms and spores are, but are not limited to, Bacillus
subtilis, Bacillus licheniformis, and Bacillus lentus. An effective
amount of the parts of, or entire broken up, microorganisms and/or
their spores is an amount sufficient to cause destruction or
deactivation of mimics, immunogens and/or antigens that cause or
exacerbate MS. In accordance with one or more such embodiments, an
effective amount of parts or entire broken up microorganisms is
from about 100 thousand CFU to about 600 billion CFU per dose,
where the dose is administered about one or more times per week, or
as often as about one to about three times daily. Methods for
breaking up suitable microorganisms and/or spores include, for
example and without limitation, sonication, crushing, shearing,
oxidation, exposure to photonic radiation or chemical (for example,
not limited to, enzymic) cleavage of the microorganisms.
[0045] In accordance with one or more embodiments of the present
invention, an above-described medicaments can be administered
orally, where an oral delivery mechanism includes, for example and
without limitation, a capsule; a tablet; a spray; and a drink,
food, powder, gum, candy, gel, or cream containing the product. The
term administered orally includes sublingually, and on an absorbent
substrate or adsorbent substrate.
[0046] In one or more embodiments of the present invention, an
above-described medicament can be administered rectally, where a
rectal delivery mechanism includes, for example and without
limitation, an enema, a fecal transplant, a gel, a cream, an
ointment, or a suppository.
[0047] In accordance with one or more embodiment, a fecal
transplant includes at least some of the following. First, a donor
of feces is screened to look for parasites, pathogenic
microorganisms, and to measure the kinds of microbes that are in
the donor's feces. The donor's feces are also analyzed for
chemicals having, for example, but not limited to, proteolytic
activity. Next, the microbial and chemical measurements are
compared against a set of requirements for a successful transplant,
for example, but not limited to, the presence of bacteria or
chemical activity that can destroy mimics, immunogens and/or
antigens that cause or exacerbate MS. Next, the donor's feces may
be corrected for pH level by adding acids, bases, or appropriate
buffering agents. Any imbalance of enzymes may be corrected by
selecting an appropriate enzyme or pro- or co-enzyme producing
microbe. A candidate microbe may be identified in the manner
described below. Next, undesirable bacteria can be neutralized or
killed. If the donor's feces do not have sufficient ability to
destroy mimics, immunogens and/or antigens that cause or exacerbate
MS, then microorganisms that are capable of destroying mimics,
immunogens and/or antigens that cause or exacerbate MS, are added,
in an effective amount, to the donor's feces prior to a fecal enema
or around the time of the fecal enema to populate the sick person's
gastrointestinal tract.
[0048] In accordance with one or more embodiments of the present
invention, enzymes for destroying or inactivating mimics,
immunogens and/or antigens are delivered to a patient. To find and
select microorganisms as sources of such enzymes, one can search
the protein database maintained by the National Institute of Health
("NIH") at http://www.ncbi.nlm.nih.gov/protein. The following is an
example of how to identify microorganisms as sources for candidate
enzymes. Enter search terms such as, for example and without
limitation, "prolyl endopeptidase," "prolyl exopeptidase," "prolyl
oligopeptidase," "subtilisin-like serine protease," and/or "prolyl
oligopeptidase-like" for enzymes that can cleave a mimic, immunogen
and/or antigen at a proline active site. The protein search engine
will return a list of microorganisms that can make such enzymes.
Identifying microorganisms as sources for enzymes such as, for
example and without limitation, a serine protease, entails entering
search terms such as, for example and without limitation, "serine
endopeptidase" and/or "serine exopeptidase" and/or "serine
oligopeptidase." Then, to make sufficient quantities of a
particular enzyme, the microorganisms identified as a result of,
but not limited to, the above-described database search can be
grown using any one of a number of methods that are well known to
those of ordinary skill in the art, and the enzyme can be extracted
from them using any one of a number of methods that are well known
to those of ordinary skill in the art. Alternatively, the relevant
gene that expresses the enzyme within the microorganism capable of
making the enzyme can be determined using any one of a number of
methods that are well known to those of ordinary skill in the art,
and the relevant gene can be inserted into another microorganism
using any one of a number of methods that are well known to those
of ordinary skill in the art to make the enzyme for extraction.
After extraction using any one of a number of methods that are well
known to those of ordinary skill in the art, and followed by
purification steps using any one of a number of methods that are
well known to those of ordinary skill in the art, if needed, the
relevant enzyme can be used in accordance with one or more
embodiments of the present invention.
[0049] In accordance with one or more embodiments, the medicament
can be administered transdermaly, where a transdermal delivery
mechanism includes, for example and without limitation, a skin
patch, a spray, a gel, a cream, an ointment or a bath.
[0050] In accordance with one or more embodiments, the medicament
can be administered intravenously, where an intravenous delivery
mechanism includes, for example and without limitation, injection
of the medicament mixed into an intravenous solution.
[0051] In accordance with one or more embodiments, the medicament
can be administered by inhalation, where an intravenous delivery
mechanism includes, for example and without limitation, a nebulized
powder inhaled by the nose or mouth.
[0052] In accordance with one or more embodiments, a medicament
comprised of subtilisin can be combined with other enzymes or
probiotics such as, for example and without limitation, PepO or
Lactobacillus lactis L1A.
[0053] Subtilisin, a serine protease or serine proteolytic enzyme,
can degrade one or more mimics of the three chemically defined MS
peptide sequences: tryptophan peptide from myelin basic protein
(see, for example, an article entitled "Essential Chemical
Requirements for Induction of Allergic Encephalomyelitis" by F. C.
Westall, A. B. Robinson, J. Caccam, J. J. Jackson and E. H. Eylar
in Nature, 229 pp. 22-24 (1971)); PHE SER TRP GLY ALA GLU GLY GLN
ARG, mid region from myelin basic protein (see, for example, an
article entitled "Biological Activity and Synthesis of an
Encephalitogenic Determinant" by R. F. Shapira, C. H. Chou, S. Mc
Kneally, E. Urban and R. F. Kibler in Science, 172 pp. 736-738
(1971)); THR THR HIS TYR GLY SER LEU PRO GLN LYS, and the
hyperacute site from myelin basic protein (see, for example, an
article entitled "Hyperacute Autoimmune Encephalomyelitis-Unique
Determinant Conferred by Serine in a Synthetic Autoantigen" by F.
C. Westall, M. Thompson and V. A. Lennon in Nature, 269 pp. 425-427
(1977)) PRO GLN LYS SER GLN ARG THR GLN ASP GLU ASN PRO VAL.
Subtilisin enzymes that are useful for treating MS are subtilisins
from any of the subtilisin proteases included in the subtilase
superfamily, family subtilisin, and subgroup true subtilisins. For
example, one or more embodiments of the present invention comprise
administering subtilisin proteases in the "true subtilisin"
subgroup which includes subtilisin BPN', subtilisin Carlsberg and
subtilisin NAT.
[0054] Serine endo- and exopeptidases are of widespread occurrence
and diverse function. Distinct families of serine proteases are
divided into six (6) superfamilies (see, for example, an article
entitled "Families of Serine Proteases" by N. D. Rawlings and A. J.
Barrett in Methods of Enzymology, 244 pp. 19-61 (1994)). The two
largest superfamilies are the chymotrypsin-like and the
subtilisin-like (subtilase) superfamilies. These two clans are
distinguished by a highly similar arrangement of catalytic His,
Asp, and Ser residues in radically different beta/beta
(chymotrypsin) and alpha/beta (subtilisin) protein scaffolds. If
pairwise sequence identity within the catalytic domains is plotted
graphically for all members of the subtilase superfamily,
clustering occurs into groups or families in which members show
higher sequence identity to each other. There are six (6) families
of which one is the "subtilisin" family. It includes mainly enzymes
from Bacillus, with subgroups of "true subtilisins" (>64%
identity), high-alkaline proteases (>55% identity), and
intracellular proteases (>37% identity) (see, for example, an
article entitled "Subtilases: The Superfamily of Subtilisin-Like
Serine Proteases" by R. J. Siezen and J. A. M. Leunissen in Protein
Sci., 6 pp. 501-523 (1997).)
[0055] In accordance with one or more embodiments of the present
invention, subtilisins used in the therapeutic treatment of MS
includes any of the subtilisin proteases included in the
superfamily subtilase, family subtilisin, and subgroup true
subtilisins. The definition the requirements of this group are set
forth in an article entitled "Subtilases: The Superfamily of
Subtilisin-Like Serine Proteases" by R. J. Siezen and J. A. M.
Leunissen in Protein Sci., 6 pp. 501-523 (1997), which article is
incorporated herein by reference in its entirety.
[0056] Myasthenia Gravis (MG): In accordance with one or more
embodiments of the present invention, a medicament comprised of an
effective dose of oligopeptidase F (PepF) is administered to a
patient to treat/cure MG. PepF belongs to the M3 metalloprotease
family. While most bacterial PepFs are cytoplasmic endopeptidases,
some are secreted, for example, the enzyme from Bacillus
amyuloliquefaciens. Pep F has been seen in a variety of bacterial
genuses including, Lactococcus and Bacillus and in Bacillus
subtilis. (see, for example, the following articles: "Biochemical
and Genetic Characterization of PepF, an Oligopeptidase from
Lactococcus Lactis" by V. Monnet, M. Nardi, A. Chopin, M.-C. Chopin
and J.-C. Gripon in J. Biol. Chem., 269 pp. 32070-32076 (1994);
"Duplication of the PepF Gene and Shuffling of DNA Fragments on the
Lactose Plasmid of Lactococcus Lactis" by M. Nardl, P. Renault and
V. Monnet in J. Bacteriol., 179 pp. 4164-4171 (1997);
"Overexpression of the PepF Oligopeptidase Inhibots Sporulation
Initiation in Bacillus Subtilis" by K. Kanamaru, K. S. Stephenson
and M. Perego in J. Bacteriol., 184 pp. 43-50 (2002); and
"Characterization of a novel Pep-F-like oligopeptidase secreted by
Bacillus amyloliquefaciens" by S.-H Chao, T.-H. Cheng, C.-Y. Shaw,
M. H. Lee, Y.-H. Hsu and Y.-C. Tsai in 23-7A. App. Environ.
Microbiol., 72 pp. 968-971 (2006)). Pep F favors breaking bonds
n-terminal to hydrophobic residues, but also can break bonds
between proline and hydrophobic residues. Furthermore, it has been
shown to break bonds n-terminal to charged amino acids. With
respect to alpha MSH, this protease should break those bonds of
peptides that mimic the c-terminal of alpha MSH. In accordance with
one or more such embodiments, an effective dose or amount of PepF
is a dose or amount of the protease (for example, in sufficient
concentration) that is effective in destroying or deactivating
mimics, immunogens antigens that cause or exacerbate MG. The
effective amount of endopeptidase F (PepF) administered will depend
upon the severity of the disease process (the medicament will be
administered one or more, preferably three, doses daily), but will
range between about 20 units/day and about 200 units/day where the
definition of a unit is an amount of PepF required to cleave one
micromole of bradykinin at a pH of 8.0 and a temperature of
40.degree. C.
[0057] In accordance with one or more further embodiments, a
medicament comprised of an effective dose of endopeptidase O2
(PepO2) is administered to a patient to treat/cure MG. In
accordance with one or more such embodiments, an effective dose or
amount of PepO2 is a dose or amount of the protease (for example,
in sufficient concentration) that is effective in destroying or
deactivating mimics, immunogens antigens that cause or exacerbate
MG. The effective amount of endopeptidase O2 (PepO2) administered
will depend upon the severity of the disease process (the
medicament will be administered one or more, preferably three,
doses daily), but will range between about 20 units/day and about
200 units/day where the definition of a unit is an amount of PepO2
required to cleave one micromole of BCN (f193-209) at a pH of 6.5
and a temperature of 25.degree. C. While endopeptidase O2 retains
an ability to break bonds n-terminal to hydrophobic residues, the
added specificity for post-proline bonds distinguishes it from
other PepO-type endopeptidases (see, for example, an article
entitled "Identification and Characterization of Lactobacillus
Helveticus PepO2, an Endopeptidase with Post-Proline Specificity"
by Y.-S. Chen, J. E. Christensen, J. R. Broadbent and J. L. Steele
in Appl. Environ. Microbiol., 69 pp. 1276-1282 (2003)).
[0058] In accordance with one or more further embodiments of the
present invention, a medicament comprised of an effective amount of
a non-pathogenic microorganism and/or its spores (that are capable
of providing PepF) is administered to a patient to treat/cure MG.
In accordance with one or more such embodiments, the microorganism
and spores include, for example and without limitation,
Lactobacillus jensenii, Lactobacillus crispatus, Lactobacillus
johnsonii, Lactobacillus plantarum, Lactobacillus helveticus,
Lactobacillus amylolyticus, Lactobacillus salivarius, Lactobacillus
ultunensis, Lactobacillus rhamnosus, Lactobacillus acidophilus,
Lactobacillus delbrueki bulgaricus, Lactobacillus gasseri,
Lactobacillus casei, Lactobacillus coleohominis, Lactobacillus
fermentum, Lactobacillus paracasei, Lactococcus lactis cremoris,
Enterococcus faecalis, Bacillus cereus (spore-forming),
Campylobacter subtilisis, and Oenococcus oeni and their various
strains. An effective amount of the microorganism and/or its spores
is an amount that is effective to cause destruction or deactivation
of mimics, immunogens and/or antigens that cause or exacerbate MG.
In accordance with one or more such embodiments, an effective
amount is from about 100 thousand CFU to about 600 billion CFU per
dose, where the dose is administered about one or more times per
week, or as often as about one to about three times daily.
[0059] In accordance with one or more embodiments of the present
invention, a medicament comprised of an effective amount of a
non-pathenogenic microorganism and/or its spores (that are capable
of providing PepO2) is administered to a patient to treat/cure MG.
In accordance with one or more such embodiments, the microorganism
and spores include, for example and without limitation, Lactococcus
lactis cremoris, Lactobacillus helveticus, and Lactobacillus
johnsonii and their various strains. An effective amount of the
microorganism and/or its spores is an amount that is effective to
cause destruction or deactivation of mimics, immunogens and/or
antigens that cause or exacerbate MG. In accordance with one or
more such embodiments, an effective amount is from about 100
thousand CFU to about 600 billion CFU per dose, where the dose is
administered about one or more times per week, or as often as about
one to three times daily.
[0060] In accordance with one or more embodiments of the present
invention, a medicament comprised of an effective amount of parts
of, or entire broken up, microorganisms and/or their spores (that
are capable of providing PepF) is administered to a patient to
treat/cure MG. Parts of the microorganisms and/or spores can be
separated and selected, using any one of a number of methods that
are well known to those of ordinary skill in the art, for their
bioactive properties to help ensure and improve the rate of the
destruction or deactivation of mimics, immunogens and/or antigens
that cause or exacerbate MG and/or to improve the effectiveness of
enzymes in the gastrointestinal or respiratory tract in destroying
or deactivating such mimics, immunogens and/or antigens. In
accordance with one or more such embodiments, the microorganism and
spores include, for example and without limitation, Lactobacillus
jensenii, Lactobacillus crispatus, Lactobacillus johnsonii,
Lactobacillus plantarum, Lactobacillus helveticus, Lactobacillus
amylolyticus, Lactobacillus salivarius, Lactobacillus ultunensis,
Lactobacillus rhamnosus, Lactobacillus acidophilus, Lactobacillus
delbrueki bulgaricus, Lactobacillus gasseri, Lactobacillus casei,
Lactobacillus coleohominis, Lactobacillus fermentum, Lactobacillus
paracasei, Lactococcus lactis cremoris, Enterococcus faecalis,
Bacillus cereus (spore-forming), and Oenococcus oeni and their
various strains. An effective amount of the parts of, or entire
broken up, microorganisms and/or their spores is an amount
sufficient to cause destruction or deactivation of mimics,
immunogens and/or antigens that cause or exacerbate MG. In
accordance with one or more such embodiments, an effective amount
of parts or entire broken up microorganisms is from about 100
thousand CFU to about 600 billion CFU per dose, where the dose is
administered about one or more times per week, or as often as about
one to three times daily. Methods for breaking up suitable
microorganisms and/or spores are set forth above with respect to
MS.
[0061] In accordance with one or more embodiments of the present
invention, a medicament comprised of an effective amount of parts
of, or entire broken up, microorganisms and/or their spores (that
are capable of providing PepO2) is administered to a patient to
treat/cure MG. Parts of the microorganisms and/or spores can be
separated and selected, using any one of a number of methods that
are well known to those of ordinary skill in the art, for their
bioactive properties to help ensure and improve the rate of the
destruction or deactivation of mimics, immunogens or immunogens
that cause or exacerbate MG and/or to improve the effectiveness of
enzymes in the gastrointestinal or respiratory tract in destroying
or deactivating such mimics, immunogens and/or antigens. In
accordance with one or more such embodiments, the microorganism and
spores include, for example and without limitation, Lactococcus
lactis cremoris, Lactobacillus helveticus, and Lactobacillus
johnsonii and their various strains. An effective amount of the
parts of, or entire broken up, microorganisms and/or their spores
is an amount sufficient to cause destruction or deactivation of
mimics, immunogens and/or antigens that cause or exacerbate MG. In
accordance with one or more such embodiments, an effective amount
of parts or entire broken up microorganisms is from about 100
thousand CFU to about 600 billion CFU per dose, where the dose is
administered about one or more times per week, or as often as about
one to three times daily. Methods for breaking up suitable
microorganisms and/or spores are set forth above with respect to
MS.
[0062] In accordance with one or more embodiments, the
above-described medicaments can be administered: orally (the
methods relating to oral administration described above with
respect to MS may also be applied with respect to MG); rectally
(the methods relating to rectal administration and selection of
microorganisms as sources of suitable enzymes described above with
respect to MS may also be applied with respect to MG);
transdermally (the methods relating to transdermal administration
described above with respect to MS may also be applied with respect
to MG); intravenously (the methods relating to intravenous
administration described above with respect to MS may also be
applied with respect to MG); and by inhalation (the methods
relating to inhalation administration described above with respect
to MS may also be applied with respect to MG).
[0063] In accordance with one or more embodiments, a medicament
comprised of PepF can be combined with other enzymes or probiotics
such as, for example and without limitation, subtilisin or
Lactobacillus lactis L1A.
[0064] Uveoretinitis: In accordance with one or more embodiments of
the present invention, a medicament comprised of an effective dose
of oligopeptidase F (PepF) is administered to a patient to
treat/cure Uveoretinitis. In accordance with one or more such
embodiments, an effective dose or amount of PepF is a dose or
amount of the protease (for example, in sufficient concentration)
that is effective in destroying or deactivating mimics, immunogens
antigens that cause or exacerbate Uveoretinitis. The effective
amount of endopeptidase F (PepF) administered will depend upon the
severity of the disease process (the medicament will be
administered one or more, preferably three, doses daily), but will
range between about 20 units/day and about 200 units/day where the
definition of a unit is an amount of PepF required to cleave one
micromole of bradykinin at a pH of 8.0 and a temperature of
40.degree. C.
[0065] In accordance with one or more further embodiments, a
medicament comprised of an effective dose of endopeptidase O (PepO)
is administered to a patient to treat/cure Uveoretinitis. This
enzyme specifically hydrolyzes 20-5 amino acids peptides on the
N-terminal side of hydrophobic amino acids (see, for example, the
following articles: "Enzymatic Ability of Bifidobacterium Animalis
Subsp Lactis to Hydrolyze Milk Proteins: Identification and
Characterization of Endopeptidase O" by C. Janer, F. Arigoni, B. H.
Lee, C, Pelaez and T. Requena in App. Environ. Microbiol., 71 pp.
8460-8465 (2005); "Cloning and expression of an oligopeptidase,
PepO, with novel specificity form Lactobacillus rhamnosus HN001" by
C. Christensson, H. Bratt, L. J. Collins. T. Coolbear, et al. in
App. Environ. Microbiol., 68 pp. 254-262 (2002); "Characterization
of an Intracellular Oligopeptidase from Lactobacillus Paracasei" by
R. O. Tobiassen, T. Sorhaug and L. Stepaniak in App. Environ.
Microbiol., 63 pp. 1284-1287 (1997); "Genetic characterization and
physiological role of endopeptidase O from Lactobacillus helveticus
CNRZ32." by Y--S Chen and J. L. Steele in App. Environ. Microbiol.,
64 pp. 3411-3415 (1998); "Cloning and Sequencing of the Gene for a
Lactococcal Endopeptidase, an Enzyme with Sequence Similarity to
Mammalian Enkephalinase" by I. Mierau, P. S. T. Tan, A. J.
Haandrikman, J. Kok et al. in J. Bacteriol., 175 pp. 2087-2096
(1993); "Purification and Characterization of an Endopeptidase from
Lactococcus Lactis Subsp. Cremoris SK11" by G. C. Pritchard, A. D.
Freebairn and T. Coolberg in Microbiol., 140 pp. 923-930 (1994);
and "Purification and Characterization of an Endopetidase from
Lactococcus Lactis Subsp Cremoris Wg2" by P. S. T. Tan, K. M. Pos
and W. N. Koninigs in App. Environ. Microbiol., 57 pp. 3593-3599
(1991)). Endopeptidase O specifically hydrolyzes 20-5 amino acids
peptides on the N-terminal side of hydrophobic amino acids, and
known immunogens fall in this range. Hence, the PepO will destroy
or help destroy mimics, immunogens and/or antigens for CD, and it
will also destroy or help destroy small biologically active proline
peptides which adversely affect Uveoretinitis patients. Thus, once
digested, the smaller peptides can be quickly absorbed and
transported into bacterial cells for further destruction to
individual amino acids.
[0066] In accordance with one or more such embodiments, an
effective dose or amount of PepO is a dose or amount of the
protease (for example, in sufficient concentration) that is
effective in destroying or deactivating mimics, immunogens antigens
that cause or exacerbate Uveoretinitis. The effective amount of
endopeptidase O (PepO) administered will depend upon the severity
of the disease process (the medicament will be administered one or
more, preferably three, doses daily), but will range between about
20 units/day and about 200 units/day where the definition of a unit
is the amount of PepO required to cleave one micromole of
bradykinin at a pH of 6.0 and a temperature of 25.degree. C.
[0067] In accordance with one or more embodiments of the present
invention, a medicament comprised of an effective amount of a
non-pathogenic microorganism and/or its spores (that are capable of
providing PepF) is administered to a patient to treat/cure
Uveoretinitis. In accordance with one or more such embodiments, the
microorganism and spores include, for example and without
limitation, Lactobacillus jensenii, Lactobacillus crispatus,
Lactobacillus johnsonii, Lactobacillus plantarum, Lactobacillus
helveticus, Lactobacillus amylolyticus, Lactobacillus salivarius,
Lactobacillus ultunensis, Lactobacillus rhamnosus, Lactobacillus
acidophilus, Lactobacillus delbrueki bulgaricus, Lactobacillus
gasseri, Lactobacillus casei, Lactobacillus coleohominis,
Lactobacillus fermentum, Lactobacillus paracasei, Lactococcus
lactis cremoris, Enterococcus faecalis, Bacillus cereus
(spore-forming), Campylobacter subtilisin, and Oenococcus oeni and
their various strains. An effective amount of the microorganism
and/or its spores is an amount that is effective to cause
destruction or deactivation of mimics, immunogens and/or antigens
that cause or exacerbate Uveoretinitis. In accordance with one or
more such embodiments, an effective amount is from about 100
thousand CFU to about 600 billion CFU per dose, where the dose is
administered about one or more times per week, or as often as about
one to about three times daily.
[0068] In accordance with one or more embodiments of the present
invention, a medicament comprised of an effective amount of a
non-pathenogenic microorganism and/or its spores (that are capable
of providing PepO) is administered to a patient to treat/cure
Uveoretinitis. Endopeptidase O is found in a large range of
bacterial systems. In accordance with one or more such embodiments,
the microorganism and spores include, for example and without
limitation, Lactobacillus sanfrancisens, Lactobacillus plantarum,
Lactobacillus acidophilus, Lactobacillus helveticus, Lactobacillus
casei, Lactobacillus reuteri, Lactobacillus salivarius,
Lactobacillus gasseri, Lactobacillus rhamnosus, Lactobacillus
johnsonii, Lactobacillus jensenii, Lactobacillus amylolyticus,
Lactobacillus sakei, Lactobacillus antrii, Lactobacillus paracasei,
Lactobacillus ruminis, Lactococcus lactis, Bifidobacterium dentium,
Bifidobacterium longum, Bifidobacterium adolescentis,
Bifidobacterium animalis, and Oenicoccus oeni and their various
strains. An effective amount of the microorganism and/or its spores
is an amount that is effective to cause destruction or deactivation
of mimics, immunogens and/or antigens that cause or exacerbate
Uveoretinitis. In accordance with one or more such embodiments, an
effective amount is from about 100 thousand CFU to about 600
billion CFU per dose, where the dose is administered about one or
more times per week, or as often as about one to three times
daily.
[0069] In accordance with one or more embodiments of the present
invention, a medicament comprised of an effective amount of parts
of, or entire broken up, microorganisms and/or their spores (that
are capable of providing PepF) is administered to a patient to
treat/cure Uveoretinitis. Parts of the microorganisms and/or spores
can be separated and selected, using any one of a number of methods
that are well known to those of ordinary skill in the art, for
their bioactive properties to help ensure and improve the rate of
the destruction or deactivation of mimics, immunogens and/or
antigens that cause or exacerbate Uveoretinitis and/or to improve
the effectiveness of enzymes in the gastrointestinal or respiratory
tract in destroying or deactivating such mimics, immunogens and/or
antigens. In accordance with one or more such embodiments, the
microorganism and spores include, for example and without
limitation, Lactobacillus jensenii, Lactobacillus crispatus,
Lactobacillus johnsonii, Lactobacillus plantarum, Lactobacillus
helveticus, Lactobacillus amylolyticus, Lactobacillus salivarius,
Lactobacillus ultunensis, Lactobacillus rhamnosus, Lactobacillus
acidophilus, Lactobacillus delbrueki bulgaricus, Lactobacillus
gasseri, Lactobacillus casei, Lactobacillus coleohominis,
Lactobacillus fermentum, Lactobacillus paracasei, Lactococcus
lactis cremoris, Enterococcus faecalis, Bacillus cereus
(spore-forming), Campylobacter subtilisin, and Oenococcus oeni and
their various strains. An effective amount of the parts of, or
entire broken up, microorganisms and/or their spores is an amount
sufficient to cause destruction or deactivation of mimics,
immunogens and/or antigens that cause or exacerbate Uveoretinitis.
In accordance with one or more such embodiments, an effective
amount of parts or entire broken up microorganisms is from about
100 thousand CFU to about 600 billion CFU per dose, where the dose
is administered about one or more times per week, or as often as
about one to three times daily. Methods for breaking up suitable
microorganisms and/or spores are set forth above with respect to
MS.
[0070] In accordance with one or more embodiments of the present
invention, a medicament comprised of an effective amount of parts
of, or entire broken up, microorganisms and/or their spores (that
are capable of providing PepO) is administered to a patient to
treat/cure Uveoretinitis. Parts of the microorganisms and/or spores
can be separated and selected, using any one of a number of methods
that are well known to those of ordinary skill in the art, for
their bioactive properties to help ensure and improve the rate of
the destruction or deactivation of mimics, immunogens or immunogens
that cause or exacerbate Uveoretinitis and/or to improve the
effectiveness of enzymes in the gastrointestinal or respiratory
tract in destroying or deactivating such mimics, immunogens and/or
antigens. In accordance with one or more such embodiments, the
microorganism and spores include, for example and without
limitation, Lactobacillus sanfrancisens, Lactobacillus plantarum,
Lactobacillus acidophilus, Lactobacillus helveticus, Lactobacillus
casei, Lactobacillus reuteri, Lactobacillus salivarius,
Lactobacillus gasseri, Lactobacillus rhamnosus, Lactobacillus
johnsonii, Lactobacillus jensenii, Lactobacillus amylolyticus,
Lactobacillus sakei, Lactobacillus antri, Lactobacillus paracasei,
Lactobacillus ruminis, Lactococcus lactis, Bifidobacterium dentium,
Bifidobacterium longum, Bifidobacterium adolescentis,
Bifidobacterium animalis, and Oenicoccus oeni and their various
strains. An effective amount of the parts of, or entire broken up,
microorganisms and/or their spores is an amount sufficient to cause
destruction or deactivation of mimics, immunogens and/or antigens
that cause or exacerbate Uveoretinitis. In accordance with one or
more such embodiments, an effective amount of parts or entire
broken up microorganisms is from about 100 thousand CFU to about
600 billion CFU per dose, where the dose is administered about one
or more times per week, or as often as about one to three times
daily. Methods for breaking up suitable microorganisms and/or
spores are set forth above with respect to MS.
[0071] In accordance with one or more embodiments, the
above-described medicaments can be administered: orally (the
methods relating to oral administration described above with
respect to MS may also be applied with respect to Uveoretinitis);
rectally (the methods relating to rectal administration and
selection of microorganisms as sources of suitable enzymes
described above with respect to MS may also be applied with respect
to Uveoretinitis); transdermally (the methods relating to
transdermal administration described above with respect to MS may
also be applied with respect to Uveoretinitis); intravenously (the
methods relating to intravenous administration described above with
respect to MS may also be applied with respect to Uveoretinitis);
and by inhalation (the methods relating to inhalation
administration described above with respect to MS may also be
applied with respect to Uveoretinitis).
[0072] In accordance with one or more embodiments, a medicament
comprised of PepF can be combined with other enzymes or probiotics
such as, for example and without limitation, subtilisin or
Lactobacillus lactis L1A.
[0073] Lupus Erythematosus: In accordance with one or more
embodiments of the present invention, a medicament comprised of an
effective dose of endopeptidase O2 (PepO2) is administered to a
patient to treat/cure Lupus Erythematosis. In accordance with one
or more such embodiments, an effective dose or amount of PepO2 is a
dose or amount of the PepO2 (for example, in sufficient
concentration) that is effective in destroying or deactivating
mimics, immunogens antigens that cause or exacerbate Lupus
Erythematosus. While endopeptidase O2 retains the ability to break
bonds n-terminal to hydrophobic residues, the added specificity for
post-proline bonds distinguishes it form other PepO-type
endopeptidases (see, for example, an article entitled
"Identification and Characterization of Lactobacillus Helveticus
PepO2, an Endopeptidase with Post-Proline Specificity" by Y.-S.
Chen, J. E. Christensen, J. R. Broadbent and J. L. Steele in Appl.
Environ. Microbiol., 69 pp. 1276-1282 (2003)). The effective amount
of endopeptidase O2 (PepO2) administered will depend upon the
severity of the disease process (the medicament will be
administered one or more, preferably three, doses daily), but will
range between about 20 units/day and about 200 units/day where the
definition of a unit is an amount of PepO2 required to cleave one
micromole of BCN (f193-209) at a pH of 6.5 and a temperature of
25.degree. C.
[0074] In accordance with one or more further embodiments, a
medicament comprised of an effective dose of endonuclease O (PepO)
from Bifidobacterium animalis subsp lactis is administered to a
patient to treat/cure Lupus Erythematosus. Such PepO will destroy
potential lupus erythematosus mimics, immunogens and/or antigens
prior to immune activation (see, for example, an article entitled
"Enzymatic Ability of Bifidobacterium Animalis Subsp Lactis to
Hydrolyze Milk Proteins: Identification and Characterization of
Endopeptidase O" by C. Janer, F. Arigoni, B. H. Lee, C, Pelaez and
T. Requena in App. Environ. Microbiol., 71 pp. 8460-8465 (2005).
While this is a PepO endonuclease, the protease from
Bifidobacterium has an ability to hydrolyze pro-pro bonds that has
not been reported with other endopeptidase O proteases.
[0075] In accordance with one or more such embodiments, an
effective dose or amount of PepO is a dose or amount of the
protease (for example, in sufficient concentration) that is
effective in destroying or deactivating mimics, immunogens antigens
that cause or exacerbate Lupus Erythematosus. The effective amount
of endopeptidase O (PepO) administered will depend upon the
severity of the disease process (the medicament will be
administered one or more, preferably three, doses daily), but will
range between about 20 units/day and about 200 units/day where the
definition of a unit is the amount of PepO from Bifdobacterium
animalis subsp. lactis required to cleave one micromole of
bradykinin at a pH of 6.0 and a temperature of 25.degree. C.
[0076] In accordance with one or more embodiments of the present
invention, a medicament comprised of an effective amount of a
non-pathenogenic microorganism and/or its spores (that are capable
of providing PepO2) is administered to a patient to treat/cure
Lupus Erythematosus. In accordance with one or more such
embodiments, the microorganism and spores include, for example and
without limitation, Lactococcus lactis cremoris, Lactobacillus
helveticus, and Lactobacillus johnsonii and their various strains.
An effective amount of the microorganism and/or its spores is an
amount that is effective to cause destruction or deactivation of
mimics, immunogens and/or antigens that cause or exacerbate Lupus
Erythematosus. In accordance with one or more such embodiments, an
effective amount is from about 100 thousand CFU to about 600
billion CFU per dose, where the dose is administered about one or
more times per week, or as often as about one to three times
daily.
[0077] In accordance with one or more embodiments of the present
invention, a medicament comprised of an effective amount of a
non-pathenogenic microorganism and/or its spores (that are capable
of providing PepO) is administered to a patient to treat/cure Lupus
Erythematosus. In accordance with one or more such embodiments, the
microorganism and spores include, for example and without
limitation, Bifidobacterium animalis subsp lactis and their various
strains. An effective amount of the microorganism and/or its spores
is an amount that is effective to cause destruction or deactivation
of mimics, immunogens and/or antigens that cause or exacerbate
Lupus Erythematosus. In accordance with one or more such
embodiments, an effective amount is from about 100 thousand CFU to
about 600 billion CFU per dose, where the dose is administered
about one or more times per week, or as often as about one to three
times daily.
[0078] In accordance with one or more embodiments of the present
invention, a medicament comprised of an effective amount of parts
of, or entire broken up, microorganisms and/or their spores (that
are capable of providing PepO2) is administered to a patient to
treat/cure Lupus Erythematosus. Parts of the microorganisms and/or
spores can be separated and selected, using any one of a number of
methods that are well known to those of ordinary skill in the art,
for their bioactive properties to help ensure and improve the rate
of the destruction or deactivation of mimics, immunogens or
immunogens that cause or exacerbate Lupus Erythematosus and/or to
improve the effectiveness of enzymes in the gastrointestinal or
respiratory tract in destroying or deactivating such mimics,
immunogens and/or antigens. In accordance with one or more such
embodiments, the microorganism and spores include, for example and
without limitation, Lactococcus lactis cremoris, Lactobacillus
helveticus, and Lactobacillus johnsonii and their various strains.
An effective amount of the parts of, or entire broken up,
microorganisms and/or their spores is an amount sufficient to cause
destruction or deactivation of mimics, immunogens and/or antigens
that cause or exacerbate Lupus Erythematosus. In accordance with
one or more such embodiments, an effective amount of parts or
entire broken up microorganisms is from about 100 thousand CFU to
about 600 billion CFU per dose, where the dose is administered
about one or more times per week, or as often as about one to three
times daily. Methods for breaking up suitable microorganisms and/or
spores are set forth above with respect to MS.
[0079] In accordance with one or more embodiments of the present
invention, a medicament comprised of an effective amount of parts
of, or entire broken up, microorganisms and/or their spores (that
are capable of providing PepO) is administered to a patient to
treat/cure Lupus Erythematosus. Parts of the microorganisms and/or
spores can be separated and selected, using any one of a number of
methods that are well known to those of ordinary skill in the art,
for their bioactive properties to help ensure and improve the rate
of the destruction or deactivation of mimics, immunogens and/or
antigens that cause or exacerbate Lupus Erythematosus and/or to
improve the effectiveness of enzymes in the gastrointestinal or
respiratory tract in destroying or deactivating such mimics,
immunogens and/or antigens. In accordance with one or more such
embodiments, the microorganism and spores include, for example and
without limitation, Bifidobacterium animalis subsp lactis and their
various strains. An effective amount of the parts of, or entire
broken up, microorganisms and/or their spores is an amount
sufficient to cause destruction or deactivation of mimics,
immunogens and/or antigens that cause or exacerbate Lupus
Erythematosus. In accordance with one or more such embodiments, an
effective amount of parts or entire broken up microorganisms is
from about 100 thousand CFU to about 600 billion CFU per dose,
where the dose is administered about one or more times per week, or
as often as about one to three times daily. Methods for breaking up
suitable microorganisms and/or spores are set forth above with
respect to MS.
[0080] In accordance with one or more embodiments, the
above-described medicaments can be administered: orally (the
methods relating to oral administration described above with
respect to MS may also be applied with respect to Lupus
Erythematosus); rectally (the methods relating to rectal
administration and selection of microorganisms as sources of
suitable enzymes described above with respect to MS may also be
applied with respect to Lupus Erythematosus); transdermally (the
methods relating to transdermal administration described above with
respect to MS may also be applied with respect to Lupus
Erythematosus); intravenously (the methods relating to intravenous
administration described above with respect to MS may also be
applied with respect to Lupus Erythematosus); and by inhalation
(the methods relating to inhalation administration described above
with respect to MS may also be applied with respect to Lupus
Erythematosus).
[0081] In accordance with one or more embodiments, a medicament
comprised of PepO or PepO2 can be combined with other enzymes or
probiotics such as, for example and without limitation, subtilisin
or Lactobacillus lactis L1A.
[0082] Ulcerative Colitis: In accordance with one or more
embodiments of the present invention, a medicament comprised of an
effective dose of oligopeptidase F (PepF) is administered to a
patient to treat/cure Ulcerative Colitis. In accordance with one or
more such embodiments, an effective dose or amount of PepF is a
dose or amount of the protease (for example, in sufficient
concentration) that is effective in destroying or deactivating
mimics, immunogens antigens that cause or exacerbate Ulcerative
Colitis. The effective amount of endopeptidase F (PepF)
administered will depend upon the severity of the disease process
(the medicament will be administered one or more, preferably three,
doses daily), but will range between about 20 units/day and about
200 units/day where the definition of a unit is an amount of PepF
required to cleave one micromole of bradykinin at a pH of 8.0 and a
temperature of 40.degree. C.
[0083] In accordance with one or more further embodiments, a
medicament comprised of an effective dose of endopeptidase O (PepO)
is administered to a patient to treat/cure Ulcerative Colitis.
Endopeptidase O is found in a large range of bacterial systems, and
specifically hydrolyzes 20-5 amino acids peptides on the N-terminal
side of hydrophobic amino acids. Hence, PepO will destroy or help
destroy mimics, immunogens and/or antigens for Ulcerative Colitis
so that they can be absorbed and transported in bacterial cells for
further destruction to individual amino acids. This process will
reduce the concentration of small proline containing peptides,
especially C-terminal proline peptides which can contribute to
hormonal aberrations. Known immunogens fall in this range. Once
digested, the smaller peptides can be quickly absorbed and
transported in the bacterial cells for further destruction to
individual amino acids (see, for example, the following articles:
"Linear Epitope Mapping of a SmB/B' Polypeptide" by J. A. James and
J. B. Harley in Immunol., 148 pp. 2074-9 (1992); "Cloning and
expression of an oligopeptidase, PepO, with novel specificity form
Lactobacillus rhamnosus HN001" by C. Christensson, H. Bratt, L. J.
Collins. T. Coolbear, et al. in App. Environ. Microbiol., 68 pp.
254-262 (2002); "Characterization of an Intracellular
Oligopeptidase from Lactobacillus Paracasei" by R. O. Tobiassen, T.
Sorhaug and L. Stepaniak in App. Environ. Microbiol., 63 pp.
1284-1287 (1997); "Genetic characterization and physiological role
of endopeptidase O from Lactobacillus helveticus CNRz32." by Y--S
Chen and J. L. Steele in App. Environ. Microbiol., 64 pp. 3411-3415
(1998); "Cloning and Sequencing of the Gene for a Lactococcal
Endopeptidase, an Enzyme with Sequence Similarity to Mammalian
Enkephaliinase" by I. Mierau, P. S. T. Tan, A. J. Haandrikman, J.
Kok et al. in J. Bacteriol., 175 pp. 2087-2096 (1993);
"Purification and Characterization of an Endopeptidase from
Lactococcus Lactis Subsp. Cremoris SK11" by G. C. Pritchard, A. D.
Freebairn and T. Coolberg in Microbiol., 140 pp. 923-930 (1994);
and "Purification and Characterization of an Endopetidase from
Lactococcus Lactis Subsp Cremoris Wg2" by P. S. T. Tan, K. M. Pos
and W. N. Koninigs in App. Environ. Microbiol., 57 pp. 3593-3599
(1991)).
[0084] In accordance with one or more such embodiments, an
effective dose or amount of PepO is a dose or amount of the
protease (for example, in sufficient concentration) that is
effective in destroying or deactivating mimics, immunogens antigens
that cause or exacerbate Ulcerative Colitis. The effective amount
of endopeptidase O (PepO) administered will depend upon the
severity of the disease process (the medicament will be
administered one or more, preferably three, doses daily), but will
range between about 20 units/day and about 200 units/day where the
definition of a unit is the amount of PepO required to cleave one
micromole of bradykinin at a pH of 6.0 and a temperature of
25.degree. C.
[0085] In accordance with one or more embodiments of the present
invention, a medicament comprised of an effective amount of a
non-pathogenic microorganism and/or its spores (that are capable of
providing PepF) is administered to a patient to treat/cure
Ulcerative Colitis. In accordance with one or more such
embodiments, the microorganism and spores include, for example and
without limitation, Lactobacillus jensenii, Lactobacillus
crispatus, Lactobacillus johnsonii, Lactobacillus plantarum,
Lactobacillus helveticus, Lactobacillus amylolyticus, Lactobacillus
salivarius, Lactobacillus ultunensis, Lactobacillus rhamnosus,
Lactobacillus acidophilus, Lactobacillus delbrueki bulgaricus,
Lactobacillus gasseri, Lactobacillus casei, Lactobacillus
coleohominis, Lactobacillus fermentum, Lactobacillus paracasei,
Lactococcus lactis cremoris, Enterococcus faecalis, Bacillus cereus
(spore-forming), and Oenococcus oeni and their various strains. An
effective amount of the microorganism and/or its spores is an
amount that is effective to cause destruction or deactivation of
mimics, immunogens and/or antigens that cause or exacerbate
Ulcerative Colitis. In accordance with one or more such
embodiments, an effective amount is from about 100 thousand CFU to
about 600 billion CFU per dose, where the dose is administered
about one or more times per week, or as often as about one to about
three times daily.
[0086] In accordance with one or more embodiments of the present
invention, a medicament comprised of an effective amount of a
non-pathenogenic microorganism and/or its spores (that are capable
of providing PepO) is administered to a patient to treat/cure
Ulcerative Colitis. In accordance with one or more such
embodiments, the microorganism and spores include, for example and
without limitation, Lactobacillus san francisens, Lactobacillus
plantarum, Lactobacillus acidophilus, Lactobacillus helveticus,
Lactobacillus casei, Lactobacillus reuteri, Lactobacillus
salivarius, Lactobacillus gasseri, Lactobacillus rhamnosus,
Lactobacillus johnsonii, Lactobacillus jensenii, Lactobacillus
amylolyticus, Lactobacillus sakei, Lactobacillus antri,
Lactobacillus paracasei, Lactobacillus ruminis, Lactococcus lactis,
Bifidobacterium dentium, Bifidobacterium longum, Bifidobacterium
adolescentis, Bifidobacterium animalis, and Oenicoccus oeni and
their various strains. An effective amount of the microorganism
and/or its spores is an amount that is effective to cause
destruction or deactivation of mimics, immunogens and/or antigens
that cause or exacerbate Ulcerative Colitis. In accordance with one
or more such embodiments, an effective amount is from about 100
thousand CFU to about 600 billion CFU per dose, where the dose is
administered about one or more times per week, or as often as about
one to three times daily.
[0087] In accordance with one or more embodiments of the present
invention, a medicament comprised of an effective amount of parts
of, or entire broken up, microorganisms and/or their spores (that
are capable of providing PepF) is administered to a patient to
treat/cure Ulcerative Colitis. Parts of the microorganisms and/or
spores can be separated and selected, using any one of a number of
methods that are well known to those of ordinary skill in the art,
for their bioactive properties to help ensure and improve the rate
of the destruction or deactivation of mimics, immunogens and/or
antigens that cause or exacerbate Ulcerative Colitis and/or to
improve the effectiveness of enzymes in the gastrointestinal or
respiratory tract in destroying or deactivating such mimics,
immunogens and/or antigens. In accordance with one or more such
embodiments, the microorganism and spores include, for example and
without limitation, Lactobacillus jensenii, Lactobacillus
crispatus, Lactobacillus johnsonii, Lactobacillus plantarum,
Lactobacillus helveticus, Lactobacillus amylolyticus, Lactobacillus
salivarius, Lactobacillus ultunensis, Lactobacillus rhamnosus,
Lactobacillus acidophilus, Lactobacillus delbrueki bulgaricus,
Lactobacillus gasseri, Lactobacillus casei, Lactobacillus
coleohominis, Lactobacillus fermentum, Lactobacillus paracasei,
Lactococcus lactis cremoris, Enterococcus faecalis, Bacillus cereus
(spore-forming), and Oenococcus oeni and their various strains. An
effective amount of the parts of, or entire broken up,
microorganisms and/or their spores is an amount sufficient to cause
destruction or deactivation of mimics, immunogens and/or antigens
that cause or exacerbate Ulcerative Colitis. In accordance with one
or more such embodiments, an effective amount of parts or entire
broken up microorganisms is from about 100 thousand CFU to about
600 billion CFU per dose, where the dose is administered about one
or more times per week, or as often as about one to three times
daily. Methods for breaking up suitable microorganisms and/or
spores are set forth above with respect to MS.
[0088] In accordance with one or more embodiments of the present
invention, a medicament comprised of an effective amount of parts
of, or entire broken up, microorganisms and/or their spores (that
are capable of providing PepO) is administered to a patient to
treat/cure Ulcerative Colitis. Parts of the microorganisms and/or
spores can be separated and selected, using any one of a number of
methods that are well known to those of ordinary skill in the art,
for their bioactive properties to help ensure and improve the rate
of the destruction or deactivation of mimics, immunogens or
immunogens that cause or exacerbate Ulcerative Colitis and/or to
improve the effectiveness of enzymes in the gastrointestinal or
respiratory tract in destroying or deactivating such mimics,
immunogens and/or antigens. In accordance with one or more such
embodiments, the microorganism and spores include, for example and
without limitation, Lactobacillus sanfrancisens, Lactobacillus
plantarum, Lactobacillus acidophilus, Lactobacillus helveticus,
Lactobacillus casei, Lactobacillus reuteri, Lactobacillus
salivarius, Lactobacillus gasseri, Lactobacillus rhamnosus,
Lactobacillus johnsonii, Lactobacillus jensenii, Lactobacillus
amylolyticus, Lactobacillus sakei, Lactobacillus antri,
Lactobacillus paracasei, Lactobacillus ruminis, Lactococcus lactis,
Bifidobacterium dentium, Bifidobacterium longum, Bifidobacterium
adolescentis, Bifidobacterium animalis, and Oenicoccus oeni and
their various strains. An effective amount of the parts of, or
entire broken up, microorganisms and/or their spores is an amount
sufficient to cause destruction or deactivation of mimics,
immunogens and/or antigens that cause or exacerbate Ulcerative
Colitis. In accordance with one or more such embodiments, an
effective amount of parts or entire broken up microorganisms is
from about 100 thousand CFU to about 600 billion CFU per dose,
where the dose is administered about one or more times per week, or
as often as about one to three times daily. Methods for breaking up
suitable microorganisms and/or spores are set forth above with
respect to MS.
[0089] In accordance with one or more embodiments, the
above-described medicaments can be administered: orally (the
methods relating to oral administration described above with
respect to MS may also be applied with respect to Ulcerative
Colitis); rectally (the methods relating to rectal administration
and selection of microorganisms as sources of suitable enzymes
described above with respect to MS may also be applied with respect
to Ulcerative Colitis); transdermally (the methods relating to
transdermal administration described above with respect to MS may
also be applied with respect to Ulcerative Colitis); intravenously
(the methods relating to intravenous administration described above
with respect to MS may also be applied with respect to Ulcerative
Colitis); and by inhalation (the methods relating to inhalation
administration described above with respect to MS may also be
applied with respect to Ulcerative Colitis).
[0090] In accordance with one or more embodiments, a medicament
comprised of PepF can be combined with other enzymes or probiotics
such as, for example and without limitation, subtilisin or
Lactobacillus lactis L1A.
[0091] Celiac Disease (CD): In accordance with one or more
embodiments of the present invention, a medicament comprised of an
effective dose or amount of Oenococcus oeni is administered to a
patient to treat/cure CD. Oenicoccus oeni is a bacterial agent used
as a starter inoculum in wine making (see, for example, the
following articles: "Saccharomyces Cerevisiae-Oenococcus Oeni
Interactions in Wine: Current Knowledge and Perspectives" by H.
Alexandre, P. J. Costello, F. Rremize, J. Guzzo et al., in Int. J.
Food Microbiol., 93 pp. 141-54 (2004); "A new approach for
selection of Oenococcus oeni strains in order to produce malolactic
starters" by F. Coucheney, N. Desroche, M. Bou, R.
Tourdot-Marechal, L. Dulau and J. Guzzo in Int. J. Food Microbiol.,
105 pp. 463-70 (2005); and "Characterization and Technological
Properties of Oenococcus Oeni Strains from Wine Spontaneous
Malolactic Fermentations: a Framework for Selection of New Starter
Cultures" by L. Solieri, F. Genova, M. De Paola and P. Giudici in
J. Appl. Microbiol., 108 pp. 285-98 (2010)). An effective dose or
amount of Oenococcus oeni is a dose or amount of Oenococcus oeni
that is effective in destroying or deactivating mimics, immunogens
and/or antigens that cause or exacerbate CD. The effective amount
of Oenococcus oeni administered will depend upon the severity of
the disease process, but each dose should contain from about
1.times.10.sup.5 to about 600.times.10.sup.9 CFU per dose of each
strain, and preferably >1.times.10.sup.7 CFU per dose of each
single strain. It is believed that Oenococcus oeni provides a
series of proline digesting enzymes which destroy small proline,
potentially harmful, peptides (see, for example, the following
articles: "Purification and Partial Characterization of Oenococcus
Oeni Exoprotease" by M. E. Farias and M. C. Manca de Nadra in FEBS
Lett., 185 pp. 263-6 (2000); "Characterization of EprA, a Major
Extracellular Protein of Oenococcus Oeni with Protease Activity" by
P. Folio, J. F. Ritt, H. Alexander and F. Remize in Int. J. Food
Microbiol., 127 pp. 26-31 (2008); and "Peptidases Specific for
Proline-Containing Peptides and Their Unusual Peptide-Dependent
Regulation in Oenococcus oeni" by J. F. Ritt, F. Remize, C.
Grandvalet, J. Guzzo, D. Atlan and H. Alexandre in J. Appl.
Microbiol., 106 pp. 801-13 (2009)). Further, Oenococcus Oeni has an
efficient transport system for 2-5 amino acid oligopeptides (see,
for example, the following articles: "Oligopeptide Assimilation and
Transport by Oenococcus Oeni" by J. F. Ritt, M. Guilloux-Benatier,
J. Guzzo, H. Alexandre and F. Remize in J. Appl. Microbiol., 104
pp. 573-80 (2008) and "Oenococcus Oeni Preference for Peptides:
Qualitative and Quantitative Analysis of Nitrogen Assimilation" by
F. Remize, A. Gaudin, Y. Kong, J. Guzzo et al. in Arch. Microbiol.,
185 pp. 459-69 (2006)). It is believed that this treatment will
help reduce the concentration of small hormonally active proteins
produced from gluten digestion from contributing negatively
biochemically, and will address a problem (see, for example, an
article entitled "The Effects of ALV003 Predigestion of Gluten on
Immune Response and Symptoms in Celiac Disease in Vivo" by J. A.
Tye-Din, R. P. Anderson, R. A. French, G. J Brown et al. in Clin.
Immunol., 134 pp. 289-295 (2010)) of clinical signs persisting and
exacerbating after gluten immunogens have been destroyed.
[0092] There are many procedures that are well known to those of
ordinary skill in the art for preparing Oenococcus oeni. The
following lists some of the more efficient methods: (a) Oenococcus
oeni strains can be cultured at 25.degree. C. in a medium adjusted
to pH 5 and containing the following (per liter) (i) yeast extract,
4 g; (ii) beef extract, 8 g; (iii) Bacto Peptone, 10 g; (iv)
glucose, 10 g; (v) fructose, 10 g; (vi) malic acid, 10 g; (vii)
KH.sub.2PO.sub.4, 2 g; (viii) MgSO.sub.4-7H.sub.2O, 0.2 g; (ix)
MnSO.sub.4--H.sub.2O, 0.1 g; and (x) Tween 80, 1 ml.; (b) strains
can be cultured at 30.degree. C. in FT80 medium (see, for example,
the following articles: "Membrane Fluidity Adjustments in Ethanol
Stressed Oenococcus Oeni cells" by M. Gracis da Silveira, E. A.
Golovina, E. Hoekstra, F. M. Rombouts and T Abee in Appl. Environ.
Microbiol., 69 pp. 5826-5832 (2003); "Medium for screening
Leuconostic Oenos Strains Defective in Malolactic Fermentation" by
J. Cavin, J. Prevost, J. Lin, P. Schmitt and C. Davies in Appl.
Environ. Microbiol., 55 pp. 751-753 (1989)) at pH 4.5 without the
addition of Tween but containing 10 g of DL-malic acid per liter
where: (i) the glucose and fructose should be autoclaved separately
and added to the medium just before inoculation at final
concentrations of 2 and 8 g/liter, and (ii) stock cultures (kept
frozen at -80.degree. C.) are to be grown until early stationary
phase (48 hr), diluted 100 fold in fresh medium, and incubated for
24 hrs.; and (c) Oenococcus oeni strains can be cultured at 30 C
and pH 5.3 in FT80 medium modified as described by an article
entitled "Lactic Acid Bacteria in the Quality Improvement and
Depreciation of Wine" by A. Louvaud-Funel in Antonie Leeuwenhoek,
76 pp. 317-331 (1999)).
[0093] In accordance with one or more further embodiments, a
medicament comprised of an effective dose of endopeptidase O (PepO)
or a combination of PepO and Oneoccocus oeni will be administered
to a patient to treat/cure CD. Endopeptidase O specifically
hydrolyzes 20-5 amino acids peptides on the N-terminal side of
hydrophobic amino acids, and known immunogens fall in this range.
Hence, the PepO will destroy or help destroy mimics, immunogens
and/or antigens for CD, and it will also destroy or help destroy
small biologically active proline peptides which adversely effect
CD patients. Thus, once digested, smaller peptides can be quickly
absorbed and transported into bacterial cells for further
destruction to individual amino acids. As a result, the
concentration of small proline containing peptides, especially
C-terminal proline peptides that can contribute to hormonal
adversities, are reduced.
[0094] In accordance with one or more such embodiments, an
effective dose or amount of PepO or a combination of PepO and
Oneococcus oeni is a dose or amount (for example, in sufficient
concentration) that is effective in destroying or deactivating
mimics, immunogens antigens that cause or exacerbate CD. The
effective amount of PepO or a combination of PepO and Oneococcus
oeni administered will depend upon the severity of the disease
process (the medicament will be administered one or more,
preferably three, doses daily), but the amount of the PepO in the
dose will range between about 20 units/day and about 200 units/day
where the definition of a unit is the amount of PepO required to
cleave one micromole of bradykinin at a pH of 6.0 and a temperature
of 25.degree. C. and the amount of the Oneococcus oeni in a dose
will range from about 1.times.10.sup.5 to about 600.times.10.sup.11
CFU per dose of each strain, and preferably >1.times.10.sup.7
CFU per dose of each single strain.
[0095] In accordance with one or more further embodiments, a
medicament comprised of an effective dose of endopeptidase O2
(PepO2) or a combination of PepO2 and Oneococcus oeni is
administered to a patient to treat/cure CD. The Lactobacillus
helveticus PepO2 enzyme contains the zinc dependent metalloprotease
motif HEXXH, and exhibits levels of amino acid homology of 72, 61,
59 and 53% to L. helveticus PepO, Lactococcus lactis PepO2,
Lactococcus lactis PepO and Lactobacillus rhamnosus PepO,
respectively. While it retains the specificity for bonds n-terminal
to hydrophobic amino acids, its added specificity for post-proline
bonds distinguishes it from other PepO-type endopeptidases (see,
for example, an article entitled "Identification and
Characterization of Lactobacillus Helveticus PepO2, an
Endopeptidase with Post-Proline Specificity" by Y.-S. Chen, J. E.
Christensen, J. R. Broadbent and J. L. Steele in Appl. Environ.
Microbiol., 69 pp. 1276-1282 (2003)).
[0096] In accordance with one or more such embodiments, an
effective dose or amount of endopeptidase O2 (PepO2) or a
combination of PepO2 and Oneococcus oeni is a dose or amount (for
example, in sufficient concentration) that is effective in
destroying or deactivating mimics, immunogens antigens that cause
or exacerbate CD. The effective amount of endopeptidase O2 (PepO2)
or a combination of PepO2 and Oneococcus oeni administered will
depend upon the severity of the disease process (the medicament
will be administered one or more, preferably three, doses daily),
and the amount of the PepO2 in the dose will range between about 20
units/day and about 200 units/day where the definition of a unit is
the amount of PepO2 required to cleave one micromole of BCN
(f193-209) at a pH of 6.5 and a temperature of 25.degree. C. and
the amount of the Oneococcus oeni in the dose will range from about
1.times.10.sup.5 to about 600.times.10.sup.9 CFU per dose of each
strain, and preferably >1.times.10.sup.7 CFU per dose of each
single strain.
[0097] In accordance with one or more further embodiments, a
medicament comprised of an effective dose of endopeptidase F (PepF)
or a combination of PepF and Oneococcus oeni is administered to a
patient to treat/cure CD. PepF, also a metallopeptidase, hydrolyzes
21-5 amino acid peptides on the N-terminal side of hydrophobic
amino acids. Its activity is similar to PepO, but it is less active
toward proline bonds.
[0098] In accordance with one or more such embodiments, an
effective dose or amount of endopeptidase F (PepF) or a combination
of PepF and Oneococcus oeni is a dose or amount (for example, in
sufficient concentration) that is effective in destroying or
deactivating mimics, immunogens antigens that cause or exacerbate
CD. The effective amount of an effective dose or amount of PepF or
a combination of PepF and Oneococcus oeni administered will depend
upon the severity of the disease process (the medicament will be
administered one or more, preferably three, doses daily), and the
amount of the PepF in the dose will range between about 20
units/day and about 200 units/day where the definition of a unit is
the amount of PepF required to cleave one micromole of bradykinin
at a pH of 8.0 and a temperature of 40.degree. C. and the amount of
the Oneococcus oeni in the dose will range from about
1.times.10.sup.5 to about 600.times.10.sup.9 CFU per dose of each
strain, and preferably >1.times.10.sup.7 CFU per dose of each
single strain.
[0099] In accordance with one or more further embodiments of the
present invention, a medicament comprised of an effective amount of
a non-pathenogenic microorganism and/or its spores (that are
capable of providing PepO) or a combination of Oneococcus oeni and
a non-pathenogenic microorganism and/or its spores (that are
capable of providing PepO) is administered to a patient to
treat/cure CD. In accordance with one or more such embodiments, the
microorganism and spores include, for example and without
limitation, Lactobacillus sanfrancisens, Lactobacillus plantarum,
Lactobacillus acidophilus, Lactobacillus helveticus, Lactobacillus
casei, Lactobacillus reuteri, Lactobacillus salivarius,
Lactobacillus gasseri, Lactobacillus rhamnosus, Lactobacillus
johnsonii, Lactobacillus jensenii, Lactobacillus amylolyticus,
Lactobacillus sakei, Lactobacillus antri, Lactobacillus paracasei,
Lactobacillus ruminis, Lactococcus lactis, Bifidobacterium dentium,
Bifidobacterium longum, Bifidobacterium adolescentis,
Bifidobacterium animalis, and Oenicoccus oeni and their various
strains. In accordance with one or more such embodiments, an
effective dose or amount of the non-pathenogenic microorganism
and/or its spores (that are capable of providing PepO) or a
combination of Oneococcus oeni and a non-pathenogenic microorganism
and/or its spores (that are capable of providing PepO) is an amount
that is effective to cause destruction or deactivation of mimics,
immunogens and/or antigens that cause or exacerbate Celiac Disease.
The effective dose administered will depend upon the severity of
the disease process (the medicament will be administered one or
more, preferably three, doses daily), but the amount of the
microorganism and spores in the dose will range from about 100
thousand CFU to about 600 billion CFU per dose, and the amount of
the Oneococcus oeni in a dose will range from about
1.times.10.sup.5 to about 600.times.10.sup.9 CFU per dose of each
strain, and preferably >1.times.10.sup.7 CFU per dose of each
single strain.
[0100] In accordance with one or more further embodiments of the
present invention, a medicament comprised of an effective amount of
a non-pathenogenic microorganism and/or its spores (that are
capable of providing PepO2) or a combination of Oenococcus oeni and
a non-pathenogenic microorganism and/or its spores (that are
capable of providing PepO2) is administered to a patient to
treat/cure CD. In accordance with one or more such embodiments, the
microorganism and spores include, for example and without
limitation, Lactococcus lactis cremoris, Lactobacillus helveticus,
and Lactobacillus johnsonii and their various strains. In
accordance with one or more such embodiments, an effective dose or
amount of the non-pathenogenic microorganism and/or its spores
(that are capable of providing PepO2) or a combination of
Oenococcus oeni and a non-pathenogenic microorganism and/or its
spores (that are capable of providing PepO2) is an amount that is
effective to cause destruction or deactivation of mimics,
immunogens and/or antigens that cause or exacerbate Celiac Disease.
The effective dose administered will depend upon the severity of
the disease process (the medicament will be administered one or
more, preferably three, doses daily), but the amount of the
microorganism and spores in the dose will range from about 100
thousand CFU to about 600 billion CFU per dose, and the amount of
the Oenococcus oeni in a dose will range from about
1.times.10.sup.5 to about 600.times.10.sup.9 CFU per dose of each
strain, and preferably >1.times.10.sup.7 CFU per dose of each
single strain.
[0101] In accordance with one or more further embodiments of the
present invention, a medicament comprised of an effective amount of
a non-pathenogenic microorganism and/or its spores (that are
capable of providing PepF) or a combination of Oenococcus oeni and
a non-pathenogenic microorganism and/or its spores (that are
capable of providing PepF) is administered to a patient to
treat/cure CD. In accordance with one or more such embodiments, the
microorganism and spores include, for example and without
limitation, but not limited to, Lactobacillus jensenii,
Lactobacillus crispatus, Lactobacillus johnsonii, Lactobacillus
plantarum, Lactobacillus helveticus, Lactobacillus amylolyticus,
Lactobacillus salivarius, Lactobacillus ultunensis, Lactobacillus
rhamnosus, Lactobacillus acidophilus, Lactobacillus delbrueki
bulgaricus, Lactobacillus gasseri, Lactobacillus casei,
Lactobacillus coleohominis, Lactobacillus fermentum, Lactobacillus
paracasei, Lactococcus lactis cremoris, Enterococcus faecalis,
Bacillus cereus (spore-forming), Campylobacter subtilisin, and
Oenococcus oeni and their various strains. In accordance with one
or more such embodiments, an effective dose or amount of the
non-pathenogenic microorganism and/or its spores (that are capable
of providing PepF) or a combination of Oenococcus oeni and a
non-pathenogenic microorganism and/or its spores (that are capable
of providing PepF) is an amount that is effective to cause
destruction or deactivation of mimics, immunogens and/or antigens
that cause or exacerbate Celiac Disease. The effective dose
administered will depend upon the severity of the disease process
(the medicament will be administered one or more, preferably three,
doses daily), but the amount of the microorganism and spores in the
dose will range from about 100 thousand CFU to about 600 billion
CFU per dose, and the amount of the Oenococcus oeni in a dose will
range from about 1.times.10.sup.5 to about 600.times.10.sup.9 CFU
per dose of each strain, and preferably >1.times.10.sup.7 CFU
per dose of each single strain.
[0102] In accordance with one or more further embodiments of the
present invention, a medicament comprised of an effective amount of
parts of, or entire broken up, microorganisms and/or their spores
(that are capable of providing PepO) or a combination of Oenococcus
oeni and parts of, or entire broken up, microorganisms and/or their
spores (that are capable of providing PepO) is administered to a
patient to treat/cure CD. Parts of the microorganisms and/or spores
can be separated and selected, using any one of a number of methods
that are well known to those of ordinary skill in the art, for
their bioactive properties to help ensure and improve the rate of
the destruction or deactivation of mimics, immunogens and/or
antigens that cause or exacerbate CD and/or to improve the
effectiveness of enzymes in the gastrointestinal or respiratory
tract in destroying or deactivating such mimics, immunogens and/or
antigens. In accordance with one or more such embodiments, the
microorganism and spores include, for example and without
limitation, Lactobacillus sanfrancisens, Lactobacillus plantarum,
Lactobacillus acidophilus, Lactobacillus helveticus, Lactobacillus
casei, Lactobacillus reuteri, Lactobacillus salivarius,
Lactobacillus gasseri, Lactobacillus rhamnosus, Lactobacillus
johnsonii, Lactobacillus jensenii, Lactobacillus amylolyticus,
Lactobacillus sakei, Lactobacillus antri, Lactobacillus paracasei,
Lactobacillus ruminis, Lactococcus lactis, Bifidobacterium dentium,
Bifidobacterium longum, Bifidobacterium adolescentis,
Bifidobacterium animalis, and Oenicoccus oeni and their various
strains. In accordance with one or more such embodiments, an
effective dose or amount of parts of, or entire broken up,
microorganisms and/or their spores (that are capable of providing
PepO) or a combination of Oenococcus oeni and parts of, or entire
broken up, microorganisms and/or their spores (that are capable of
providing PepO) is an amount that is effective to cause destruction
or deactivation of mimics, immunogens and/or antigens that cause or
exacerbate Celiac Disease. The effective dose administered will
depend upon the severity of the disease process (the medicament
will be administered one or more, preferably three, doses daily),
but the amount of parts of, or entire broken up, microorganisms
and/or their spores will range from about 100 thousand CFU to about
600 billion CFU per dose, and the amount of the Oenococcus oeni in
a dose will range from about 1.times.10.sup.5 to about
600.times.10.sup.9 CFU per dose of each strain, and preferably
>1.times.10.sup.7 CFU per dose of each single strain. Methods
for breaking up suitable microorganisms and/or spores are set forth
above with respect to MS.
[0103] In accordance with one or more further embodiments of the
present invention, a medicament comprised of an effective amount of
parts of, or entire broken up, microorganisms and/or their spores
(that are capable of providing PepO2) or a combination of
Oenococcus oeni and parts of, or entire broken up, microorganisms
and/or their spores (that are capable of providing PepO2) is
administered to a patient to treat/cure CD. Parts of the
microorganisms and/or spores can be separated and selected, using
any one of a number of methods that are well known to those of
ordinary skill in the art, for their bioactive properties to help
ensure and improve the rate of the destruction or deactivation of
mimics, immunogens and/or antigens that cause or exacerbate CD
and/or to improve the effectiveness of enzymes in the
gastrointestinal or respiratory tract in destroying or deactivating
such mimics, immunogens and/or antigens. In accordance with one or
more such embodiments, the microorganism and spores include, for
example and without limitation, Lactococcus lactis cremoris,
Lactobacillus helveticus, and Lactobacillus johnsonii and their
various strains. In accordance with one or more such embodiments,
an effective dose or amount of parts of, or entire broken up,
microorganisms and/or their spores (that are capable of providing
PepO2) or a combination of Oenococcus oeni and parts of, or entire
broken up, microorganisms and/or their spores (that are capable of
providing PepO2) is an amount that is effective to cause
destruction or deactivation of mimics, immunogens and/or antigens
that cause or exacerbate Celiac Disease. The effective dose
administered will depend upon the severity of the disease process
(the medicament will be administered one or more, preferably three,
doses daily), but the amount of parts of, or entire broken up,
microorganisms and/or their spores will range from about 100
thousand CFU to about 600 billion CFU per dose, and the amount of
the Oenococcus oeni in a dose will range from about
1.times.10.sup.5 to about 600.times.10.sup.9 CFU per dose of each
strain, and preferably >1.times.10.sup.7 CFU per dose of each
single strain. Methods for breaking up suitable microorganisms
and/or spores are set forth above with respect to MS.
[0104] In accordance with one or more further embodiments of the
present invention, a medicament comprised of an effective amount of
parts of, or entire broken up, microorganisms and/or their spores
(that are capable of providing PepF) or a combination of Oenococcus
oeni and parts of, or entire broken up, microorganisms and/or their
spores (that are capable of providing PepF) is administered to a
patient to treat/cure CD. Parts of the microorganisms and/or spores
can be separated and selected, using any one of a number of methods
that are well known to those of ordinary skill in the art, for
their bioactive properties to help ensure and improve the rate of
the destruction or deactivation of mimics, immunogens and/or
antigens that cause or exacerbate CD and/or to improve the
effectiveness of enzymes in the gastrointestinal or respiratory
tract in destroying or deactivating such mimics, immunogens and/or
antigens. In accordance with one or more such embodiments, the
microorganism and spores include, for example and without
limitation, Lactobacillus jensenii, Lactobacillus crispatus,
Lactobacillus johnsonii, Lactobacillus plantarum, Lactobacillus
helveticus, Lactobacillus amylolyticus, Lactobacillus salivarius,
Lactobacillus ultunensis, Lactobacillus rhamnosus, Lactobacillus
acidophilus, Lactobacillus delbrueki bulgaricus, Lactobacillus
gasseri, Lactobacillus casei, Lactobacillus coleohominis,
Lactobacillus fermentum, Lactobacillus paracasei, Lactococcus
lactis cremoris, Enterococcus faecalis, Bacillus cereus
(spore-forming), and Oenococcus oeni and their various strains. In
accordance with one or more such embodiments, an effective dose or
amount of parts of, or entire broken up, microorganisms and/or
their spores (that are capable of providing PepF) or a combination
of Oenococcus oeni and parts of, or entire broken up,
microorganisms and/or their spores (that are capable of providing
PepF) is an amount that is effective to cause destruction or
deactivation of mimics, immunogens and/or antigens that cause or
exacerbate Celiac Disease. The effective dose administered will
depend upon the severity of the disease process (the medicament
will be administered one or more, preferably three, doses daily),
but the amount of parts of, or entire broken up, microorganisms
and/or their spores will range from about 100 thousand CFU to about
600 billion CFU per dose, and the amount of the Oenococcus oeni in
a dose will range from about 1.times.10.sup.5 to about
600.times.10.sup.9 CFU per dose of each strain, and preferably
>1.times.10.sup.7 CFU per dose of each single strain. Methods
for breaking up suitable microorganisms and/or spores are set forth
above with respect to MS.
[0105] In accordance with one or more embodiments, the
above-described medicaments can be administered: orally (the
methods relating to oral administration described above with
respect to MS may also be applied with respect to CD); rectally
(the methods relating to rectal administration and selection of
microorganisms as sources of suitable enzymes described above with
respect to MS may also be applied with respect to CD);
transdermally (the methods relating to transdermal administration
described above with respect to MS may also be applied with respect
to CD); intravenously (the methods relating to intravenous
administration described above with respect to MS may also be
applied with respect to CD); and by inhalation (the methods
relating to inhalation administration described above with respect
to MS may also be applied with respect to CD).
[0106] Anorexia Nervosa: In accordance with one or more embodiments
of the present invention, a medicament comprised of an effective
dose of oligopeptidase F (PepF) is administered to a patient to
treat/cure Anorexia Nervosa. In accordance with one or more such
embodiments, an effective dose or amount of PepF is a dose or
amount of the protease (for example, in sufficient concentration)
that is effective in destroying or deactivating mimics, immunogens
antigens that cause or exacerbate Anorexia Nervosa. The effective
amount of endopeptidase F (PepF) administered will depend upon the
severity of the disease process (the medicament will be
administered one or more, preferably three, doses daily), but will
range between about 20 units/day and about 200 units/day where the
definition of a unit is an amount of PepF required to cleave one
micromole of bradykinin at a pH of 8.0 and a temperature of
40.degree. C.
[0107] In accordance with one or more further embodiments of the
present invention, a medicament comprised of an effective dose or
amount of subtilisin is administered to a patient to treat/cure
Anorexia Nervosa. An effective dose or amount of subtilisin is a
dose or amount of the protease that is effective in destroying or
deactivating mimics, immunogens and/or antigens that cause or
exacerbate Anorexia Nervosa. The effective amount of subtilisin
administered will depend upon the severity of the disease process,
but will range between about 2,000 fibrinolytic units/day and about
10,000 fibrinolytic units/day. For example, the subtilisin will be
administered in one or more, preferably three, doses daily of
subtilisin. It is believed that hydrolytic processes of the
subtilisin will act in two areas: the gut and the blood. In the
gut, the subtilisin will destroy or deactivate potential Anorexia
Nervosa mimics before they can reach a sufficient level for immune
activation, and, in the blood, the subtilisin will destroy any
Anorexia Nervosa immunogens and antigens that could interact with
the immune system and prolong a clinical exacerbation.
[0108] Subtilisin enzymes that are useful for treating Anorexia
Nervosa are subtilisins from any of the subtilisin proteases
included in the subtilase superfamily, family subtilisin, and
subgroup true subtilisins. For example, one or more embodiments of
the present invention comprise administering subtilisin proteases
in the "true subtilisin" subgroup which includes subtilisin BPN',
subtilisin Carlsberg and subtilisin NAT.
[0109] In accordance with one or more further embodiments of the
present invention, a medicament comprised of an effective amount of
a non-pathogenic microorganism and/or its spores (that are capable
of providing PepF) is administered to a patient to treat/cure
Anorexia Nervosa. In accordance with one or more such embodiments,
the microorganism and spores include, for example and without
limitation, Lactobacillus jensenii, Lactobacillus crispatus,
Lactobacillus johnsonii, Lactobacillus plantarum, Lactobacillus
helveticus, Lactobacillus amylolyticus, Lactobacillus salivarius,
Lactobacillus ultunensis, Lactobacillus rhamnosus, Lactobacillus
acidophilus, Lactobacillus delbrueki bulgaricus, Lactobacillus
gasseri, Lactobacillus casei, Lactobacillus coleohominis,
Lactobacillus fermentum, Lactobacillus paracasei, Lactococcus
lactis cremoris, Enterococcus faecalis, Bacillus cereus
(spore-forming), Campylobacter subtilisin, and Oenococcus oeni and
their various strains. In accordance with one or more such
embodiments, an effective dose or amount of the non-pathenogenic
microorganism and/or its spores (that are capable of providing
PepF) is an amount that is effective to cause destruction or
deactivation of mimics, immunogens and/or antigens that cause or
exacerbate Anorexia Nervosa. The effective dose administered will
depend upon the severity of the disease process (the medicament
will be administered one or more, preferably three, doses daily),
but the amount of the microorganism and spores in the dose will
range from about 100 thousand CFU to about 600 billion CFU per
dose, where the dose is administered about one or more times per
week, or as often as about one to about three times daily.
[0110] In accordance with one or more further embodiments of the
present invention, a medicament comprised of an effective amount of
a non-pathogenic microorganism and/or its spores (that are capable
of providing subtilisin) is administered to a patient to treat/cure
Anorexia Nervosa. In accordance with one or more such embodiments,
the microorganism and spores include, for example and without
limitation, Bacillus subtilis, Bacillus licheniformis, and Bacillus
lentus and their various strains. In accordance with one or more
such embodiments, an effective dose or amount of the
non-pathenogenic microorganism and/or its spores (that are capable
of providing subtilisin) is an amount that is effective to cause
destruction or deactivation of mimics, immunogens and/or antigens
that cause or exacerbate Anorexia Nervosa. The effective dose
administered will depend upon the severity of the disease process
(the medicament will be administered one or more, preferably three,
doses daily), but the amount of the microorganism and spores in the
dose will range from about 100 thousand CFU to about 600 billion
CFU per dose.
[0111] In accordance with one or more further embodiments of the
present invention, a medicament comprised of an effective amount of
parts of, or entire broken up, microorganisms and/or their spores
(that are capable of providing PepF) is administered to a patient
to treat/cure Anorexia Nervosa. Parts of the microorganisms and/or
spores can be separated and selected, using any one of a number of
methods that are well known to those of ordinary skill in the art,
for their bioactive properties to help ensure and improve the rate
of the destruction or deactivation of mimics, immunogens and/or
antigens that cause or exacerbate Anorexia Nervosa and/or to
improve the effectiveness of enzymes in the gastrointestinal or
respiratory tract in destroying or deactivating such mimics,
immunogens and/or antigens. In accordance with one or more such
embodiments, the microorganism and spores include, for example and
without limitation, Lactobacillus jensenii, Lactobacillus
crispatus, Lactobacillus johnsonii, Lactobacillus plantarum,
Lactobacillus helveticus, Lactobacillus amylolyticus, Lactobacillus
salivarius, Lactobacillus ultunensis, Lactobacillus rhamnosus,
Lactobacillus acidophilus, Lactobacillus delbrueki bulgaricus,
Lactobacillus gasseri, Lactobacillus casei, Lactobacillus
coleohominis, Lactobacillus fermentum, Lactobacillus paracasei,
Lactococcus lactis cremoris, Enterococcus faecalis, Bacillus cereus
(spore-forming), Campylobacter subtilisin, and Oenococcus oeni and
their various strains. In accordance with one or more such
embodiments, an effective dose or amount of parts of, or entire
broken up, microorganisms and/or their spores (that are capable of
providing PepF) is an amount that is effective to cause destruction
or deactivation of mimics, immunogens and/or antigens that cause or
exacerbate Anorexia Nervosa. The effective dose administered will
depend upon the severity of the disease process (the medicament
will be administered one or more, preferably three, doses daily),
but the amount of parts of, or entire broken up, microorganisms
and/or their spores will range from about 100 thousand CFU to about
600 billion CFU per dose. Methods for breaking up suitable
microorganisms and/or spores are set forth above with respect to
MS.
[0112] In accordance with one or more further embodiments of the
present invention, a medicament comprised of an effective amount of
parts of, or entire broken up, microorganisms and/or their spores
(that are capable of providing subtilisin) is administered to a
patient to treat/cure Anorexia Nervosa. Parts of the microorganisms
and/or spores can be separated and selected, using any one of a
number of methods that are well known to those of ordinary skill in
the art, for their bioactive properties to help ensure and improve
the rate of the destruction or deactivation of mimics, immunogens
and/or antigens that cause or exacerbate Anorexia Nervosa and/or to
improve the effectiveness of enzymes in the gastrointestinal or
respiratory tract in destroying or deactivating such mimics,
immunogens and/or antigens. In accordance with one or more such
embodiments, the microorganism and spores include, for example and
without limitation, Bacillus subtilis, Bacillus licheniformis, and
Bacillus lentus and their various strains. In accordance with one
or more such embodiments, an effective dose or amount of parts of,
or entire broken up, microorganisms and/or their spores (that are
capable of providing subtilisin) is an amount that is effective to
cause destruction or deactivation of mimics, immunogens and/or
antigens that cause or exacerbate Anorexia Nervosa. The effective
dose administered will depend upon the severity of the disease
process (the medicament will be administered one or more,
preferably three, doses daily), but the amount of parts of, or
entire broken up, microorganisms and/or their spores will range
from about 100 thousand CFU to about 600 billion CFU per dose.
Methods for breaking up suitable microorganisms and/or spores are
set forth above with respect to MS.
[0113] In accordance with one or more embodiments, the
above-described medicaments can be administered: orally (the
methods relating to oral administration described above with
respect to MS may also be applied with respect to Anorexia
Nervosa); rectally (the methods relating to rectal administration
and selection of microorganisms as sources of suitable enzymes
described above with respect to MS may also be applied with respect
to Anorexia Nervosa); transdermally (the methods relating to
transdermal administration described above with respect to MS may
also be applied with respect to Anorexia Nervosa); intravenously
(the methods relating to intravenous administration described above
with respect to MS may also be applied with respect to Anorexia
Nervosa); and by inhalation (the methods relating to inhalation
administration described above with respect to MS may also be
applied with respect to Anorexia Nervosa).
[0114] In accordance with one or more embodiments, a medicament
comprised of PepF can be combined with other enzymes or probiotics
such as, for example and without limitation, subtilisin or
Lactobacillus lactis L1A.
[0115] Rheumatoid Arthritis: In accordance with one or more further
embodiments of the present invention, a medicament comprised of an
effective dose or amount of subtilisin is administered to a patient
to treat/cure Rheumatoid Arthritis. An effective dose or amount of
subtilisin is a dose or amount of the protease that is effective in
destroying or deactivating mimics, immunogens and/or antigens that
cause or exacerbate Rheumatoid Arthritis. The effective amount of
subtilisin administered will depend upon the severity of the
disease process, but will range between about 2,000 fibrinolytic
units/day and about 10,000 fibrinolytic units/day. For example, the
subtilisin will be administered in one or more, preferably three,
doses daily of subtilisin. It is believed that hydrolytic processes
of the subtilisin will act in two areas: the gut and the blood. In
the gut, the subtilisin will destroy or deactivate potential
Rheumatoid Arthritis mimics before they can reach a sufficient
level for immune activation, and, in the blood, the subtilisin will
destroy or deactivate Rheumatoid Arthritis immunogens and antigens
released from cartilage that could interact with the immune system
and prolong a clinical exacerbation.
[0116] Subtilisin enzymes that are useful for treating Anorexia
Nervosa are subtilisins from any of the subtilisin proteases
included in the subtilase superfamily, family subtilisin, and
subgroup true subtilisins. For example, one or more embodiments of
the present invention comprise administering subtilisin proteases
in the "true subtilisin" subgroup which includes subtilisin BPN',
subtilisin Carlsberg and subtilisin NAT.
[0117] In accordance with one or more further embodiments, a
medicament comprised of an effective dose of endonuclease O (PepO)
from Bifidobacterium animalis subsp lactis is administered to a
patient to treat/cure Rheumatoid Arthritis. While this is a PepO
endonuclease, the protease from Bifidobacterium has an ability to
hydrolyze pro-pro bonds that has not been reported with other
endopeptidase O proteases.
[0118] In accordance with one or more such embodiments, an
effective dose or amount of PepO is a dose or amount of the
protease (for example, in sufficient concentration) that is
effective in destroying or deactivating mimics, immunogens antigens
that cause or exacerbate Rheumatoid Arthritis. The effective amount
of endopeptidase O (PepO) administered will depend upon the
severity of the disease process (the medicament will be
administered one or more, preferably three, doses daily), but will
range between about 20 units/day and about 200 units/day where the
definition of a unit is the amount of PepO from Bifdobacterium
animalis subsp. lactis required to cleave one micromole of
bradykinin at a pH of 6.0 and a temperature of 25.degree. C.
[0119] In accordance with one or more further embodiments of the
present invention, a medicament comprised of an effective dose of
endopeptidase O2 (PepO2) is administered to a patient to treat/cure
Rheumatoid Arthritis. In accordance with one or more such
embodiments, an effective dose or amount of PepO2 is a dose or
amount of the PepO2 (for example, in sufficient concentration) that
is effective in destroying or deactivating mimics, immunogens
antigens that cause or exacerbate Rheumatoid Arthritis. The
effective amount of endopeptidase O2 (PepO2) administered will
depend upon the severity of the disease process (the medicament
will be administered one or more, preferably three, doses daily),
but will range between about 20 units/day and about 200 units/day
where the definition of a unit is an amount of PepO2 required to
cleave one micromole of BCN (f193-209) at a pH of 6.5 and a
temperature of 25.degree. C.
[0120] In accordance with one or more further embodiments of the
present invention, a medicament comprised of an effective amount of
a non-pathogenic microorganism and/or its spores (that are capable
of providing subtilisin) is administered to a patient to treat/cure
Rheumatoid Arthritis. In accordance with one or more such
embodiments, the microorganism and spores include, for example and
without limitation, Bacillus subtilis, Bacillus licheniformis, and
Bacillus lentus and their various strains. In accordance with one
or more such embodiments, an effective dose or amount of the
non-pathenogenic microorganism and/or its spores (that are capable
of providing subtilisin) is an amount that is effective to cause
destruction or deactivation of mimics, immunogens and/or antigens
that cause or exacerbate Rheumatoid Arthritis. The effective dose
administered will depend upon the severity of the disease process
(the medicament will be administered one or more, preferably three,
doses daily), but the amount of the microorganism and spores in the
dose will range from about 100 thousand CFU to about 600 billion
CFU per dose.
[0121] In accordance with one or more further embodiments of the
present invention, a medicament comprised of an effective amount of
a non-pathogenic microorganism and/or its spores (that are capable
of providing PepO) is administered to a patient to treat/cure
Rheumatoid Arthritis. In accordance with one or more such
embodiments, the microorganism and spores include, for example and
without limitation, Bifidobacterium animalis subsp lactis and their
various strains. In accordance with one or more such embodiments,
an effective dose or amount of the non-pathenogenic microorganism
and/or its spores (that are capable of providing PepO) is an amount
that is effective to cause destruction or deactivation of mimics,
immunogens and/or antigens that cause or exacerbate Rheumatoid
Arthritis. The effective dose administered will depend upon the
severity of the disease process (the medicament will be
administered one or more, preferably three, doses daily), but the
amount of the microorganism and spores in the dose will range from
about 100 thousand CFU to about 600 billion CFU per dose, where the
dose is administered about one or more times per week, or as often
as about one to about three times daily.
[0122] In accordance with one or more further embodiments of the
present invention, a medicament comprised of an effective amount of
a non-pathogenic microorganism and/or its spores (that are capable
of providing PepO2) is administered to a patient to treat/cure
Rheumatoid Arthritis. In accordance with one or more such
embodiments, the microorganism and spores include, for example and
without limitation, Lactococcus lactis cremoris, Lactobacillus
helveticus, and Lactobacillus johnsonii and their various strains.
In accordance with one or more such embodiments, an effective dose
or amount of the non-pathenogenic microorganism and/or its spores
(that are capable of providing PepO2) is an amount that is
effective to cause destruction or deactivation of mimics,
immunogens and/or antigens that cause or exacerbate Rheumatoid
Arthritis. The effective dose administered will depend upon the
severity of the disease process (the medicament will be
administered one or more, preferably three, doses daily), but the
amount of the microorganism and spores in the dose will range from
about 100 thousand CFU to about 600 billion CFU per dose, where the
dose is administered about one or more times per week, or as often
as about one to about three times daily.
[0123] In accordance with one or more further embodiments of the
present invention, a medicament comprised of an effective amount of
parts of, or entire broken up, microorganisms and/or their spores
(that are capable of providing subtilisin) is administered to a
patient to treat/cure Rheumatoid Arthritis. Parts of the
microorganisms and/or spores can be separated and selected, using
any one of a number of methods that are well known to those of
ordinary skill in the art, for their bioactive properties to help
ensure and improve the rate of the destruction or deactivation of
mimics, immunogens and/or antigens that cause or exacerbate
Rheumatoid Arthritis and/or to improve the effectiveness of enzymes
in the gastrointestinal or respiratory tract in destroying or
deactivating such mimics, immunogens and/or antigens. In accordance
with one or more such embodiments, the microorganism and spores
include, for example and without limitation, Bacillus subtilis,
Bacillus licheniformis, and Bacillus lentus. In accordance with one
or more such embodiments, an effective dose or amount of parts of,
or entire broken up, microorganisms and/or their spores (that are
capable of providing subtilisin) is an amount that is effective to
cause destruction or deactivation of mimics, immunogens and/or
antigens that cause or exacerbate Rheumatoid Arthritis. The
effective dose administered will depend upon the severity of the
disease process (the medicament will be administered one or more,
preferably three, doses daily), but the amount of parts of, or
entire broken up, microorganisms and/or their spores will range
from about 100 thousand CFU to about 600 billion CFU per dose.
Methods for breaking up suitable microorganisms and/or spores are
set forth above with respect to MS.
[0124] In accordance with one or more further embodiments of the
present invention, a medicament comprised of an effective amount of
parts of, or entire broken up, microorganisms and/or their spores
(that are capable of providing PepO) is administered to a patient
to treat/cure Rheumatoid Arthritis. Parts of the microorganisms
and/or spores can be separated and selected, using any one of a
number of methods that are well known to those of ordinary skill in
the art, for their bioactive properties to help ensure and improve
the rate of the destruction or deactivation of mimics, immunogens
and/or antigens that cause or exacerbate Rheumatoid Arthritis
and/or to improve the effectiveness of enzymes in the
gastrointestinal or respiratory tract in destroying or deactivating
such mimics, immunogens and/or antigens. In accordance with one or
more such embodiments, the microorganism and spores include, for
example and without limitation, Bifidobacterium animalis subsp
lactis and their various strains. In accordance with one or more
such embodiments, an effective dose or amount of parts of, or
entire broken up, microorganisms and/or their spores (that are
capable of providing PepO) is an amount that is effective to cause
destruction or deactivation of mimics, immunogens and/or antigens
that cause or exacerbate Rheumatoid Arthritis. The effective dose
administered will depend upon the severity of the disease process
(the medicament will be administered one or more, preferably three,
doses daily), but the amount of parts of, or entire broken up,
microorganisms and/or their spores will range from about 100
thousand CFU to about 600 billion CFU per dose. Methods for
breaking up suitable microorganisms and/or spores are set forth
above with respect to MS.
[0125] In accordance with one or more further embodiments of the
present invention, a medicament comprised of an effective amount of
parts of, or entire broken up, microorganisms and/or their spores
(that are capable of providing PepO2) is administered to a patient
to treat/cure Rheumatoid Arthritis. Parts of the microorganisms
and/or spores can be separated and selected, using any one of a
number of methods that are well known to those of ordinary skill in
the art, for their bioactive properties to help ensure and improve
the rate of the destruction or deactivation of mimics, immunogens
and/or antigens that cause or exacerbate Rheumatoid Arthritis
and/or to improve the effectiveness of enzymes in the
gastrointestinal or respiratory tract in destroying or deactivating
such mimics, immunogens and/or antigens. In accordance with one or
more such embodiments, the microorganism and spores include, for
example and without limitation, Lactococcus lactis cremoris,
Lactobacillus helveticus, and Lactobacillus johnsonii and their
various strains. In accordance with one or more such embodiments,
an effective dose or amount of parts of, or entire broken up,
microorganisms and/or their spores (that are capable of providing
PepO) is an amount that is effective to cause destruction or
deactivation of mimics, immunogens and/or antigens that cause or
exacerbate Rheumatoid Arthritis. The effective dose administered
will depend upon the severity of the disease process (the
medicament will be administered one or more, preferably three,
doses daily), but the amount of parts of, or entire broken up,
microorganisms and/or their spores will range from about 100
thousand CFU to about 600 billion CFU per dose. Methods for
breaking up suitable microorganisms and/or spores are set forth
above with respect to MS.
[0126] In accordance with one or more embodiments, the
above-described medicaments can be administered: orally (the
methods relating to oral administration described above with
respect to MS may also be applied with respect to Rheumatoid
Arthritis); rectally (the methods relating to rectal administration
and selection of microorganisms as sources of suitable enzymes
described above with respect to MS may also be applied with respect
to Rheumatoid Arthritis); transdermally (the methods relating to
transdermal administration described above with respect to MS may
also be applied with respect to Rheumatoid Arthritis);
intravenously (the methods relating to intravenous administration
described above with respect to MS may also be applied with respect
to Rheumatoid Arthritis); and by inhalation (the methods relating
to inhalation administration described above with respect to MS may
also be applied with respect to Rheumatoid Arthritis).
[0127] In accordance with one or more embodiments, a medicament
comprised of PepF can be combined with other enzymes or probiotics,
and examples of other enzymes or probiotics such as, for example
and without limitation, subtilisin or Lactobacillus lactis L1A.
[0128] Autistic Spectrum Disorder: Autistic Spectrum Disorder
patients have gluten and casein sensitivities, molecules that are
high in concentration of proline, and some autistic children have
been found with antibodies to myelin basic protein. Thus, in
accordance with one or more embodiments of the present invention, a
medicament comprised of an effective dose or amount of Oenococcus
oeni is administered to a patient to treat/cure Autistic Spectrum
Disorder (referred to herein as Autism). It is believed that this
treatment will help reduce the concentration of small hormonally
active proteins produced from gluten digestion from contributing
negatively biochemically, and will address a problem of clinical
signs persisting and exacerbating after gluten immunogens have been
destroyed. An effective dose or amount of Oenococcus oeni is a dose
or amount of Oenococcus oeni that is effective in destroying or
deactivating mimics, immunogens and/or antigens that cause or
exacerbate Autistic Spectrum Disorder. The effective amount of
Oenococcus oeni administered will depend upon the severity of the
disease process, but each dose should contain from about
1.times.10.sup.5 to about 600.times.10.sup.9 CFU per dose of each
strain, and preferably >1.times.10.sup.7 CFU per dose of each
single strain.
[0129] In accordance with one or more further embodiments, a
medicament comprised of an effective dose of endopeptidase O (PepO)
or a combination of PepO and Oenococcus oeni will be administered
to a patient to treat/cure Autistic Spectrum Disorder. PepO
specifically hydrolyzes 20-5 amino acids peptides on the N-terminal
side of hydrophobic amino acids, and known immunogens fall in this
range. Hence, the PepO will destroy or help destroy mimics,
immunogens and/or antigens for Autistic Spectrum Disorder, and it
will also destroy or help destroy small biologically active proline
peptides which adversely affect Autism patients. Thus, once
digested, smaller peptides can be quickly absorbed and transported
into bacterial cells for further destruction to individual amino
acids. As a result, the concentration of small proline containing
peptides, especially C-terminal proline peptides that can
contribute to hormonal adversities, are reduced.
[0130] In accordance with one or more such embodiments, an
effective dose or amount of PepO or a combination of PepO and
Oenococcus oeni is a dose or amount (for example, in sufficient
concentration) that is effective in destroying or deactivating
mimics, immunogens antigens that cause or exacerbate Autistic
Spectrum Disorder. The effective amount of PepO or a combination of
PepO and Oenococcus oeni administered will depend upon the severity
of the disease process (the medicament will be administered one or
more, preferably three, doses daily), but the amount of the PepO in
the dose will range between about 20 units/day and about 200
units/day where the definition of a unit is the amount of PepO
required to cleave one micromole of bradykinin at a pH of 6.0 and a
temperature of 25.degree. C. and the amount of the Oenococcus oeni
in a dose will range from about 1.times.10.sup.5 to about
600.times.10.sup.9 CFU per dose of each strain, and preferably
>1.times.10.sup.7 CFU per dose of each single strain.
[0131] In accordance with one or more further embodiments, a
medicament comprised of an effective dose of endopeptidase O2
(PepO2) or a combination of PepO2 and Oenococcus oeni is
administered to a patient to treat/cure Autistic Spectrum Disorder.
In accordance with one or more such embodiments, an effective dose
or amount of endopeptidase O2 (PepO2) or a combination of PepO2 and
Oenococcus oeni is a dose or amount (for example, in sufficient
concentration) that is effective in destroying or deactivating
mimics, immunogens antigens that cause or exacerbate Autistic
Spectrum Disorder. The effective amount of endopeptidase O2 (PepO2)
or a combination of PepO2 and Oenococcus oeni administered will
depend upon the severity of the disease process (the medicament
will be administered one or more, preferably three, doses daily),
and the amount of the PepO2 in the dose will range between about 20
units/day and about 200 units/day where the definition of a unit is
the amount of PepO2 required to cleave one micromole of BCN
(f193-209) at a pH of 6.5 and a temperature of 25.degree. C. and
the amount of the Oenococcus oeni in the dose will range from about
1.times.10.sup.5 to about 600.times.10.sup.9 CFU per dose of each
strain, and preferably >1.times.10.sup.7 CFU per dose of each
single strain.
[0132] In accordance with one or more further embodiments, a
medicament comprised of an effective dose of endopeptidase F (PepF)
or a combination of PepF and Oenococcus oeni is administered to a
patient to treat/cure Autistic Spectrum Disorder. This enzyme, also
a metallopeptidase, hydrolyzes 21-5 (for Bacillus amyloliquefaciens
PepF; see, for example, an article entitled "Characterization of a
novel Pep-F-like oligopeptidase secreted by Bacillus
amyloliquefaciens" by S.-H Chao, T.-H. Cheng, C.-Y. Shaw, M. H.
Lee, Y.-H. Hsu and Y.-C. Tsai in 23-7A. App. Environ. Microbiol.,
72 pp. 968-971 (2006)) and 17-7 (for Lactococcus lactis; see, for
example, an article entitled "Biochemical and Genetic
Characterization of PepF, an Oligopeptidase from Lactococcus
Lactis" by V. Monnet, M. Nardi, A. Chopin, M.-C. Chopin and J.-C.
Gripon in J. Biol. Chem., 269 pp. 32070-32076 (1994)) amino acid
peptides on the N-terminal side of hydrophobic amino acids. Its
activity is similar to PepO, but it is less active toward proline
bonds. The enzyme has also been reported in Bacillus subtilis.
[0133] In accordance with one or more such embodiments, an
effective dose or amount of endopeptidase F (PepF) or a combination
of PepF and Oenococcus oeni is a dose or amount (for example, in
sufficient concentration) that is effective in destroying or
deactivating mimics, immunogens antigens that cause or exacerbate
Autistic Spectrum Disorder. The effective amount of an effective
dose or amount of PepF or a combination of PepF and Oenococcus oeni
administered will depend upon the severity of the disease process
(the medicament will be administered one or more, preferably three,
doses daily), and the amount of the PepF in the dose will range
between about 20 units/day and about 200 units/day where the
definition of a unit is the amount of PepF required to cleave one
micromole of bradykinin at a pH of 8.0 and a temperature of
40.degree. C. and the amount of the Oenococcus oeni in the dose
will range from about 1.times.10.sup.5 to about 600.times.10.sup.9
CFU per dose of each strain, and preferably >1.times.10.sup.7
CFU per dose of each single strain.
[0134] In accordance with one or more further embodiments of the
present invention, a medicament comprised of an effective amount of
a non-pathenogenic microorganism and/or its spores (that are
capable of providing PepO) or a combination of Oenococcus oeni and
a non-pathenogenic microorganism and/or its spores (that are
capable of providing PepO) is administered to a patient to
treat/cure Autistic Spectrum Disorder. In accordance with one or
more such embodiments, the microorganism and spores include, for
example and without limitation, Lactobacillus sanfrancisens,
Lactobacillus plantarum, Lactobacillus acidophilus, Lactobacillus
helveticus, Lactobacillus casei, Lactobacillus reuteri,
Lactobacillus salivarius, Lactobacillus gasseri, Lactobacillus
rhamnosus, Lactobacillus johnsonii, Lactobacillus jensenii,
Lactobacillus amylolyticus, Lactobacillus sakei, Lactobacillus
antri, Lactobacillus paracasei, Lactobacillus ruminis, Lactococcus
lactis, Bifidobacterium dentium, Bifidobacterium longum,
Bifidobacterium adolescentis, Bifidobacterium animalis, and
Oenicoccus oeni and their various strains. In accordance with one
or more such embodiments, an effective dose or amount of the
non-pathenogenic microorganism and/or its spores (that are capable
of providing PepO) or a combination of Oenococcus oeni and a
non-pathenogenic microorganism and/or its spores (that are capable
of providing PepO) is an amount that is effective to cause
destruction or deactivation of mimics, immunogens and/or antigens
that cause or exacerbate Autistic Spectrum Disorder. The effective
dose administered will depend upon the severity of the disease
process (the medicament will be administered one or more,
preferably three, doses daily), but the amount of the microorganism
and spores in the dose will range from about 100 thousand CFU to
about 600 billion CFU per dose, and the amount of the Oenococcus
oeni in a dose will range from about 1.times.10.sup.5 to about
600.times.10.sup.9 CFU per dose of each strain, and preferably
>1.times.10.sup.7 CFU per dose of each single strain.
[0135] In accordance with one or more further embodiments of the
present invention, a medicament comprised of an effective amount of
a non-pathenogenic microorganism and/or its spores (that are
capable of providing PepO2) or a combination of Oenococcus oeni and
a non-pathenogenic microorganism and/or its spores (that are
capable of providing PepO2) is administered to a patient to
treat/cure Autistic Spectrum Disorder. In accordance with one or
more such embodiments, the microorganism and spores include, for
example and without limitation, Lactococcus lactis cremoris,
Lactobacillus helveticus, and Lactobacillus johnsonii and their
various strains. In accordance with one or more such embodiments,
an effective dose or amount of the non-pathenogenic microorganism
and/or its spores (that are capable of providing PepO2) or a
combination of Oenococcus oeni and a non-pathenogenic microorganism
and/or its spores (that are capable of providing PepO2) is an
amount that is effective to cause destruction or deactivation of
mimics, immunogens and/or antigens that cause or exacerbate
Autistic Spectrum Disorder. The effective dose administered will
depend upon the severity of the disease process (the medicament
will be administered one or more, preferably three, doses daily),
but the amount of the microorganism and spores in the dose will
range from about 100 thousand CFU to about 600 billion CFU per
dose, and the amount of the Oenococcus oeni in a dose will range
from about 1.times.10.sup.5 to about 600.times.10.sup.9 CFU per
dose of each strain, and preferably >1.times.10.sup.7 CFU per
dose of each single strain.
[0136] In accordance with one or more further embodiments of the
present invention, a medicament comprised of an effective amount of
a non-pathenogenic microorganism and/or its spores (that are
capable of providing PepF) or a combination of Oenococcus oeni and
a non-pathenogenic microorganism and/or its spores (that are
capable of providing PepF) is administered to a patient to
treat/cure Autistic Spectrum Disorder. In accordance with one or
more such embodiments, the microorganism and spores include, for
example and without limitation, but not limited to, Lactobacillus
jensenii, Lactobacillus crispatus, Lactobacillus johnsonii,
Lactobacillus plantarum, Lactobacillus helveticus, Lactobacillus
amylolyticus, Lactobacillus salivarius, Lactobacillus ultunensis,
Lactobacillus rhamnosus, Lactobacillus acidophilus, Lactobacillus
delbrueki bulgaricus, Lactobacillus gasseri, Lactobacillus casei,
Lactobacillus coleohominis, Lactobacillus fermentum, Lactobacillus
paracasei, Lactococcus lactis cremoris, Enterococcus faecalis,
Bacillus cereus (spore-forming), Campylobacter subtilisin, and
Oenococcus oeni and their various strains. In accordance with one
or more such embodiments, an effective dose or amount of the
non-pathenogenic microorganism and/or its spores (that are capable
of providing PepF) or a combination of Oenococcus oeni and a
non-pathenogenic microorganism and/or its spores (that are capable
of providing PepF) is an amount that is effective to cause
destruction or deactivation of mimics, immunogens and/or antigens
that cause or exacerbate Autistic Spectrum Disorder. The effective
dose administered will depend upon the severity of the disease
process (the medicament will be administered one or more,
preferably three, doses daily), but the amount of the microorganism
and spores in the dose will range from about 100 thousand CFU to
about 600 billion CFU per dose, and the amount of the Oenococcus
oeni in a dose will range from about 1.times.10.sup.5 to about
600.times.10.sup.9 CFU per dose of each strain, and preferably
>1.times.10.sup.7 CFU per dose of each single strain.
[0137] In accordance with one or more further embodiments of the
present invention, a medicament comprised of an effective amount of
parts of, or entire broken up, microorganisms and/or their spores
(that are capable of providing PepO) or a combination of Oenococcus
oeni parts of, or entire broken up, microorganisms and/or their
spores (that are capable of providing PepO) is administered to a
patient to treat/cure Autistic Spectrum Disorder. Parts of the
microorganisms and/or spores can be separated and selected, using
any one of a number of methods that are well known to those of
ordinary skill in the art, for their bioactive properties to help
ensure and improve the rate of the destruction or deactivation of
mimics, immunogens and/or antigens that cause or exacerbate
Autistic Spectrum Disorder and/or to improve the effectiveness of
enzymes in the gastrointestinal or respiratory tract in destroying
or deactivating such mimics, immunogens and/or antigens. In
accordance with one or more embodiments of this invention, the
parts of the microorganisms and/or spores chosen can be, but are
not limited to, adjuvants such as muramyl dipeptide. The adjuvant's
content can be increased or decreased to saturate or starve the
gastrointestinal tract or respiratory tract of the adjuvant to
prevent the optimum ratio of adjuvant to mimic from being close
enough to one to initiate and/or maintain the autoimmune reaction.
In accordance with one or more such embodiments, the microorganism
and spores include, for example and without limitation,
Lactobacillus sanfrancisens, Lactobacillus plantarum, Lactobacillus
acidophilus, Lactobacillus helveticus, Lactobacillus casei,
Lactobacillus reuteri, Lactobacillus salivarius, Lactobacillus
gasseri, Lactobacillus rhamnosus, Lactobacillus johnsonii,
Lactobacillus jensenii, Lactobacillus amylolyticus, Lactobacillus
sakei, Lactobacillus antri, Lactobacillus paracasei, Lactobacillus
ruminis, Lactococcus lactis, Bifidobacterium dentium,
Bifidobacterium longum, Bifidobacterium adolescentis,
Bifidobacterium animalis, and Oenicoccus oeni and their various
strains. In accordance with one or more such embodiments, an
effective dose or amount of parts of, or entire broken up,
microorganisms and/or their spores (that are capable of providing
PepO) or a combination of Oenococcus oeni and parts of, or entire
broken up, microorganisms and/or their spores (that are capable of
providing PepO) is an amount that is effective to cause destruction
or deactivation of mimics, immunogens and/or antigens that cause or
exacerbate Autistic Spectrum Disorder. The effective dose
administered will depend upon the severity of the disease process
(the medicament will be administered one or more, preferably three,
doses daily), but the amount of parts of, or entire broken up,
microorganisms and/or their spores will range from about 100
thousand CFU to about 600 billion CFU per dose, and the amount of
the Oenococcus oeni in a dose will range from about
1.times.10.sup.5 to about 600.times.10.sup.9 CFU per dose of each
strain, and preferably >1.times.10.sup.7 CFU per dose of each
single strain. Methods for breaking up suitable microorganisms
and/or spores are set forth above with respect to MS.
[0138] In accordance with one or more further embodiments of the
present invention, a medicament comprised of an effective amount of
parts of, or entire broken up, microorganisms and/or their spores
(that are capable of providing PepO2) or a combination of
Oenococcus oeni and parts of, or entire broken up, microorganisms
and/or their spores (that are capable of providing PepO2) is
administered to a patient to treat/cure Autistic Spectrum Disorder.
Parts of the microorganisms and/or spores can be separated and
selected, using any one of a number of methods that are well known
to those of ordinary skill in the art, for their bioactive
properties to help ensure and improve the rate of the destruction
or deactivation of mimics, immunogens and/or antigens that cause or
exacerbate Autistic Spectrum Disorder and/or to improve the
effectiveness of enzymes in the gastrointestinal or respiratory
tract in destroying or deactivating such mimics, immunogens and/or
antigens. In accordance with one or more such embodiments, the
microorganism and spores include, for example and without
limitation, Lactococcus lactis cremoris, Lactobacillus helveticus,
and Lactobacillus johnsonii and their various strains. In
accordance with one or more such embodiments, an effective dose or
amount of parts of, or entire broken up, microorganisms and/or
their spores (that are capable of providing PepO2) or a combination
of Oenocooccus oeni and parts of, or entire broken up,
microorganisms and/or their spores (that are capable of providing
PepO2) is an amount that is effective to cause destruction or
deactivation of mimics, immunogens and/or antigens that cause or
exacerbate Autistic Spectrum Disorder. The effective dose
administered will depend upon the severity of the disease process
(the medicament will be administered one or more, preferably three,
doses daily), but the amount of parts of, or entire broken up,
microorganisms and/or their spores will range from about 100
thousand CFU to about 600 billion CFU per dose, and the amount of
the Oenococcus oeni in a dose will range from about
1.times.10.sup.5 to about 600.times.10.sup.9 CFU per dose of each
strain, and preferably >1.times.10.sup.7 CFU per dose of each
single strain. Methods for breaking up suitable microorganisms
and/or spores are set forth above with respect to MS.
[0139] In accordance with one or more further embodiments of the
present invention, a medicament comprised of an effective amount of
parts of, or entire broken up, microorganisms and/or their spores
(that are capable of providing PepF) or a combination of Oenococcus
oeni and parts of, or entire broken up, microorganisms and/or their
spores (that are capable of providing PepF) is administered to a
patient to treat/cure Autistic Spectrum Disorder. Parts of the
microorganisms and/or spores can be separated and selected, using
any one of a number of methods that are well known to those of
ordinary skill in the art, for their bioactive properties to help
ensure and improve the rate of the destruction or deactivation of
mimics, immunogens and/or antigens that cause or exacerbate
Autistic Spectrum Disorder and/or to improve the effectiveness of
enzymes in the gastrointestinal or respiratory tract in destroying
or deactivating such mimics, immunogens and/or antigens. In
accordance with one or more such embodiments, the microorganism and
spores include, for example and without limitation, Lactobacillus
jensenii, Lactobacillus crispatus, Lactobacillus johnsonii,
Lactobacillus plantarum, Lactobacillus helveticus, Lactobacillus
amylolyticus, Lactobacillus salivarius, Lactobacillus ultunensis,
Lactobacillus rhamnosus, Lactobacillus acidophilus, Lactobacillus
delbrueki bulgaricus, Lactobacillus gasseri, Lactobacillus casei,
Lactobacillus coleohominis, Lactobacillus fermentum, Lactobacillus
paracasei, Lactococcus lactis cremoris, Enterococcus faecalis,
Bacillus cereus (spore-forming), and Oenococcus oeni and their
various strains. In accordance with one or more such embodiments,
an effective dose or amount of parts of, or entire broken up,
microorganisms and/or their spores (that are capable of providing
PepF) or a combination of Oenococcus oeni and parts of, or entire
broken up, microorganisms and/or their spores (that are capable of
providing PepF) is an amount that is effective to cause destruction
or deactivation of mimics, immunogens and/or antigens that cause or
exacerbate Autistic Spectrum Disorder. The effective dose
administered will depend upon the severity of the disease process
(the medicament will be administered one or more, preferably three,
doses daily), but the amount of parts of, or entire broken up,
microorganisms and/or their spores will range from about 100
thousand CFU to about 600 billion CFU per dose, and the amount of
the Oenococcus oeni in a dose will range from about
1.times.10.sup.5 to about 600.times.10.sup.9 CFU per dose of each
strain, and preferably >1.times.10.sup.7 CFU per dose of each
single strain. Methods for breaking up suitable microorganisms
and/or spores are set forth above with respect to MS.
[0140] In accordance with one or more further embodiments of the
present invention, a medicament comprised of an effective dose or
amount of subtilisin is administered to a patient to treat/cure
Autistic Spectrum Disorder. An effective dose or amount of
subtilisin is a dose or amount of the protease that is effective in
destroying or deactivating mimics, immunogens and/or antigens that
cause or exacerbate Autistic Spectrum Disorder. The effective
amount of subtilisin administered will depend upon the severity of
the disease process, but will range between about 2,000
fibrinolytic units/day and about 10,000 fibrinolytic units/day. For
example, the subtilisin will be administered in one or more,
preferably three, doses daily of subtilisin. It is believed that
hydrolytic processes of the subtilisin will act in two areas: the
gut and the blood. In the gut, the subtilisin will destroy or
deactivate potential Autistic Spectrum Disorder mimics before they
can reach a sufficient level for immune activation, and, in the
blood, the subtilisin will destroy any Autistic Spectrum Disorder
immunogens and antigens that could interact with the immune system
and prolong a clinical exacerbation.
[0141] Subtilisin enzymes that are useful for treating Autistic
Spectrum Disorder are subtilisins from any of the subtilisin
proteases included in the subtilase superfamily, family subtilisin,
and subgroup true subtilisins. For example, one or more embodiments
of the present invention comprise administering subtilisin
proteases in the "true subtilisin" subgroup which includes
subtilisin BPN', subtilisin Carlsberg and subtilisin NAT.
[0142] In accordance with one or more further embodiments of the
present invention, a medicament comprised of an effective amount of
a non-pathogenic microorganism and/or its spores (that are capable
of providing subtilisin) is administered to a patient to treat/cure
Autistic Spectrum Disorder. In accordance with one or more such
embodiments, the microorganism and spores include, for example and
without limitation, Bacillus subtilis, Bacillus licheniformis, and
Bacillus lentus and their various strains. In accordance with one
or more such embodiments, an effective dose or amount of the
non-pathenogenic microorganism and/or its spores (that are capable
of providing subtilisin) is an amount that is effective to cause
destruction or deactivation of mimics, immunogens and/or antigens
that cause or exacerbate Autistic Spectrum Disorder. The effective
dose administered will depend upon the severity of the disease
process (the medicament will be administered one or more,
preferably three, doses daily), but the amount of the microorganism
and spores in the dose will range from about 100 thousand CFU to
about 600 billion CFU per dose.
[0143] In accordance with one or more further embodiments of the
present invention, a medicament comprised of an effective amount of
parts of, or entire broken up, microorganisms and/or their spores
(that are capable of providing subtilisin) is administered to a
patient to treat/cure Autistic Spectrum Disorder. Parts of the
microorganisms and/or spores can be separated and selected, using
any one of a number of methods that are well known to those of
ordinary skill in the art, for their bioactive properties to help
ensure and improve the rate of the destruction or deactivation of
mimics, immunogens and/or antigens that cause or exacerbate
Autistic Spectrum Disorder and/or to improve the effectiveness of
enzymes in the gastrointestinal or respiratory tract in destroying
or deactivating such mimics, immunogens and/or antigens. In
accordance with one or more such embodiments, the microorganism and
spores include, for example and without limitation, Bacillus
subtilis, Bacillus licheniformis, and Bacillus lentus and their
various strains. In accordance with one or more such embodiments,
an effective dose or amount of parts of, or entire broken up,
microorganisms and/or their spores (that are capable of providing
subtilisin) is an amount that is effective to cause destruction or
deactivation of mimics, immunogens and/or antigens that cause or
exacerbate Autistic Spectrum Disorder. The effective dose
administered will depend upon the severity of the disease process
(the medicament will be administered one or more, preferably three,
doses daily), but the amount of parts of, or entire broken up,
microorganisms and/or their spores will range from about 100
thousand CFU to about 600 billion CFU per dose. Methods for
breaking up suitable microorganisms and/or spores are set forth
above with respect to MS.
[0144] In accordance with one or more embodiments, the
above-described medicaments can be administered: orally (the
methods relating to oral administration described above with
respect to MS may also be applied with respect to Autistic Spectrum
Disorder); rectally (the methods relating to rectal administration
and selection of microorganisms as sources of suitable enzymes
described above with respect to MS may also be applied with respect
to Autistic Spectrum Disorder); transdermally (the methods relating
to transdermal administration described above with respect to MS
may also be applied with respect to Autistic Spectrum Disorder);
intravenously (the methods relating to intravenous administration
described above with respect to MS may also be applied with respect
to Autistic Spectrum Disorder); and by inhalation (the methods
relating to inhalation administration described above with respect
to MS may also be applied with respect to Autistic Spectrum
Disorder).
[0145] The following sets forth, for example and without
limitation, methods for preparing useful probiotic microorganisms.
Fermentation: As an example, microorganism Bacillus subtilis Natto
produces the endoprotease subtilisin. Fermentation additives may be
added to a culture of the microorganisms to enhance: production of
microorganisms, ability of the microorganisms to survive in the
gastrointestinal tract, ability of the microorganisms to adhere to
the gastrointestinal tract, ability of the microorganisms to
secrete desired proteases, ability of the microorganisms to secrete
chemicals to enhance survival of proteases, ability of the
microorganisms to secrete chemicals to enhance effectiveness of
desired proteases, and ability of the microorganisms to secrete
chemicals to interfere with undesired chemicals. Also, the amount
and kinds of sugars, vitamins, amino acids, proteins and/or fats
available to the microorganisms, prior to drying and forming a
powder, affect their viability. Examples of useful sugars are, but
are not limited to, sucrose, fructose, glucose, lactose, trehalose,
raffinose, paliainose, lactulose, lactitol, xylitol, sorbitol,
mannitol, malstose, dextrin and maltodextrin. Examples of useful
anti-oxidants are, but are not limited to, ascorbic acid,
glutathione and alpha-lipoic acid. Examples of useful amino acids
or their salts are, but are not limited to, lysine, cysteine,
glycine and glutamate. Examples of useful oils are, but are not
limited to, butter, palm oil, nut oil, cocoa oil, rapeseed oil and
soy bean oil. Examples of useful stabilizing ingredients are, but
are not limited to, soybean oligosaccharides,
frutooligosaccharides, galactooligosaccharides, galactosyl lactose,
milk, milk powders, whey, whey protein concentrates, casein, casein
hydrolysates, lactoferrin, lactoperoxidase, lactoglobulins,
glymacropeptides, lacto-saccharides, glycomacropeptides,
lacto-saccharides and lacto-lipids.
[0146] A chemical that inactivates an enzyme is, for example and
without limitation, a serpin. Thus, it is desirable to inhibit
serpins that inactivate proteases that destroy mimics, antigens, or
immunogens that cause autoimmune disease. Also, protective agents
such as, for example and without limitation, cryoprotectants or
other chemicals such as gels, starches, polysaccharides, and/or
sugars can be added to the culture to protect the microorganisms
during the manufacturing processes.
[0147] Removing Liquid: When a powdered form of a probiotic is
required, the microorganisms need to have liquid removed. One
option, known to those of ordinary skill in the art, is to
centrifuge the fermentation mixture to reduce the amount of liquid.
Other options include, but are not limited to, settling and
membrane filtration. This concentrates the microbes by separating
them from their supernatant. As such, it allows for more microbes
per kilogram of dried product to keep transportation and storage
costs down and to deliver more microbes per capsule or pill. It
also allows the microbes to be added as a concentrated powder to be
mixed into a drink or sprinkled onto food. Centrifuging is
beneficial when the supernatant does not contain substantial
amounts of bioactive substances that the microbes cannot readily
secrete or create in the gastrointestinal tract. However, if the
supernatant has the bioactive substances for therapeutic effect,
then centrifuging does not need to be performed if the microbes are
still desired. In some cases, centrifuging or other separation
processes, known to those of ordinary skill in the art, may be
desired to obtain the bioactive substances, such as enzymes or
bacteriocins, for delivery without the microbe. The following
methods may also be used to separate bugs from supernatant:
sedimentation, ultrafiltration and reverse osmosis.
[0148] Drying: This step dries the microorganisms as well as any
available supernatant. The microorganisms can be dried with
freeze-drying techniques, known to those of ordinary skill in the
art, by placing the centrifuged microbes and residual supernatant,
which form a slurry, onto trays and freezing them in a vacuum
environment. After the slurry dries, it resembles a cake. The dried
cake is then crushed, and the crushed powders are sieved to obtain
the desired particle size distribution. This process of drying in
bulk followed by crushing often kills many bacteria due to the
thermal and mechanical stresses applied to the microbes. Another
method of making a powder, known by those of ordinary skill in the
art, is to spray dry the microorganisms. For this process the
slurry is sprayed through a nozzle into a heated air environment.
The incoming slurry can be heated or unheated. If the shear forces
and temperatures that the microorganisms and/or enzymes experience
during the heated spray drying process are too great,
microorganisms will die or be damaged enough that the intended
therapeutic effectiveness of the microorganism and/or enzyme will
be diminished. To improve the yield and prevent damage to surviving
microorganisms, an electrospray drying process can be used.
Examples of manufacturers who make suitable electrospray drying
equipment are Charge Injection Technology and Zoom Essence.
[0149] Blending: Other ingredients, such as but not limited to,
dried proteases, microorganisms, protective sugars,
polysaccharides, gums, oils, desiccants, anti-oxidants, and
bacteriocins, can be added prior to or after the drying process.
These ingredients will assist the microbes in surviving during
storage as well as in passing to the target areas of the
gastrointestinal tract. Also adding other ingredients is needed to
reduce the dosage of concentrated microbial powders to the dosages
required for delivery to the person.
[0150] Delivery: A powder can also be an acceptable delivery system
especially when microbes do not need to be alive and where either
their microbial parts or their secreted bioactive substances are
effective against pathogens or for normalizing the protease ratios.
The powder can be consumed by adding the powder to a food or drink
product. The powder containing the microorganisms and/or enzymes
can be formulated by those of ordinary skill in the art into a
drink that may contain for example, but not limited to, water,
sweeteners, flavorings, colorants, anti-oxidants, vitamins,
minerals, short-chain fatty acids, stimulants, mood-enhancers,
teas, anti-inflammatories, and other bioactive ingredients. The
powder can be consumed after sprinkling or pouring it over solid
food or mixing into a liquid. The powder can be packaged into bulk
containers such as a bag or can or into individual sachets for easy
of carrying and single use dosing. To form a tablet, a powder
containing the microbes and/or enzyme(s), excipients, and/or other
bioactive substances are compressed into a mold in a tableting
machine. The tablet can be coated with methods and processes known
to those of ordinary skill in the art to prevent the premature
dissolution of the product in the stomach to keep the microbes
alive for delivery further down the gastrointestinal (GI) tract.
Such coatings are designed by those of ordinary skill in the art to
dissolve by time in the GI tract or more preferably by pH exposure
as the pH along the GI tract is acidic in the stomach and the pH
increases by the time the digested contents reach the large
intestine. At the large intestine, the pH is approximately 7. Some
examples known to those of ordinary skill in the art of a pH
triggered coating are, but not limited to, Eudragit and shellac.
The tablet can be designed by those of ordinary skill in the art to
be consumed orally or inserted into the rectum or vagina as a
suppository. To form a capsule, a powder containing the
microorganisms, enzymes, excipients and/or other bioactive
substances are directed into a capsule that can be made of
materials known to those of ordinary skill in the art, but are not
limited to, hardened gelatin or other polymer. The capsule can be
coated by processes known to those of ordinary skill in the art to
prevent the premature dissolution of the product in the stomach to
keep the microorganisms alive and enzymes effective for delivery
further down the GI tract. Such coatings known to those who are of
ordinary skill in the art are designed to dissolve by time in the
GI tract or more preferably by pH exposure. Some examples known to
those of ordinary skill in the art of a pH triggered coating are,
but not limited to, Eudragit and shellac. The capsule can be
designed by those of ordinary skill in the art to be consumed
orally or inserted into the rectum or vagina as a suppository. An
alternate form of a capsule to contain the microorganisms, enzymes,
excipients, and/or other bioactive substances is a gel capsule that
can be made of materials and processes known to those of ordinary
skill in the art.
[0151] For a liquid delivery system, the microorganisms and/or
enzymes and bioactive substances can be introduced simply in a
fermented liquid. That liquid can be in the form of cultured or
non-cultured animal-based and/or plant-based milk such as, but not
limited to, cow's, goat's, rice, almond, and/or soy milk.
Alternatively, microorganisms and/or enzymes can added to a drink
such, as but not limited to, a juice or formulated into a drink
that may contain for example but not limited to water, sweeteners,
flavorings, colorants, anti-oxidants, vitamins, minerals,
short-chain fatty acids, stimulants, mood-enhancers, teas,
anti-inflammatories and other bioactive ingredients. For a solid
delivery system the microorganisms and/or enzymes and bioactive
substances can be added to solid food in accordance with a number
of methods that are well known to those of ordinary skill in the
art. Examples of such food are, but are not limited to, candy,
confectionary, chewing gum, energy bars, fermented/dried
vegetables, fermented/dried meat, fermented/dried seafood,
fermented/dried fruit, fermented/dried beans and frozen desserts.
For a slurry delivery system the microorganisms and/or enzymes and
bioactive substances can be added to slurry foods in accordance
with a number of methods that are well known to those of ordinary
skill in the art. Examples of such food are, but not limited to,
yogurt, jams, jellies, gravies, gel shots, puddings, frozen
desserts, salad dressings, syrups and spreads.
[0152] Embodiments of the present invention described above are
exemplary, and many changes and modifications may be made to the
description set forth above by those of ordinary skill in the art
while remaining within the scope of the invention. For example, the
amino acid sequence of each peptidase discussed above will vary
with each strain of the bacterial types discussed above. However,
the mere changing of the amino acid sequence does not necessarily
alter its biological properties and eliminate it from a particular
proteolytic class. As such, the scope of the invention should be
determined with reference to the appended claims along with their
full scope of equivalents.
TABLE-US-00001 APPENDIX I Table 1. Chemically Defined Autoimmune
Disease-Producing Immunogens Description Sequence a. Tryptophan
peptide from myelin basic PHE SER TRP GLY protein (EAE) (see an
article entitled ALA GLU GLY GLN "Essential Chemical Requirements
ARG for Induction of Allergic Encephalomyelitis" by F. C. Westall,
A. B. Robinson, J. Caccam, J. J. Jackson and E. H. Eylar in Nature,
229 pp. 22-24 (1971)) b. Mid region from myelin basic THR THR HIS
TYR protein (EAE) (see an article GLY SER LEU PRO entitled
"Biological Activity and GLN LYS Synthesis of an Encephalitogenic
Determinant" by R. F. Shapira, C. H. Chou, S. Mc Kneally, E. Urban
and R. F. Kibler in Science, 172 pp. 736-738 (1971)) c. Hyperacute
site from myelin basic PRO GLN LYS SER protein (EAE) (see an
article GLN ARG THR GLN entitled "Hyperacute Autoimmune ASP GLU ASN
PRO Encephalomyelitis-Unique Determinant VAL Conferred by Serine in
a Synthetic Autoantigen" by F. C. Westall, M. Thompson and V. A.
Lennon in Nature, 269 pp. 425-427 (1977)) d. S-antigen 375-386 (see
an article entitled PHE VAL PHE GLU "Structure-function studies of
s-antigen: GLU PHE ALA ARG use of proteases to reveal a dominant
GLN ASN LEU LYS uveitogenic site" by H. S. Dua, P. Hossain, P. A.
J. Brown, A. McKinnon, J. V. Forrester, S. Gregerson and L. A.
Donoso in Autoimmunity, 10 pp. 153-163 (1991)) (EAU) e. S-antigen
352-364 (EAU) PRO PHE ARG LEU (see an article entitled
"Identification MET HIS PRO GLN of a Potent New Pathogenic Site in
PRO GLU ASP PRO Human Retinal S-Antigen which Induces ASP
Experimental Autoimmune Uveoretinitis in LEW Rats" by D. S.
Gregerson, C. F. Merryman, W. F. Obritsch and L. A. Donoso in Cell.
Immunol., 128 pp. 209-219 (1990)) f. Interphotoreceptor retinoid
TYR ILE ILE SER binding protein (IRBP)165-177 (EAU) TYR LEU HIS PRO
(see an article entitled "Identification of a GLY ASN THR ILE Major
Pathogenic Epitope in the Human LEU IRBP Molecule Recognized by
Mice of the H-2 Hapotype" by P. B. Silver, L. V. Rizzo, C.-C. Chan,
L. A. Donoso, B. Wiggert and R. R. Caspi in Invest. Ophthalmol.
Vis. Sci., 36 pp. 946-954 (1995)) g. Interphotoreceptor retinoid
binding PHE GLN PRO SER protein 5-20 (see an article entitled LEU
VAL LEU ASP "Identification of a new epitope of MET ALA LYS VAL
human IRBP that induces LEU LEU ASP autoimmune uveoretinitis in
mice of the H-2b haplotype" by D. Avichezer, P. B. Silver, C. -C.
Chan, B. Wiggert and R. R. Caspi in Invest. Opthalmol. Vis. Sci.,
41 pp. 127-131 (2000)) (EAU) h. Interphotoreceptor retinoid binding
ALA ASP LYS ASP protein 201-216 (EAU) (see an article VAL VAL LEU
THR entitled "Identification of a Peptide SER SER ARG THR Inducing
Experimental Autoimmune GLY GLY VAL Uveoretinitis in H2Ak-Carrying
Mice" by K. Namba, K. Ogasawara, N. Kitaichi, A. Matsuki et al. in
Clin. Exp. Immunol., 111 pp. 442-449 (1998). i. Acetylcholine
receptor 129-145 (EMG) GLU ILE ILE VAL (see an article entitled "A
17-Mer Self- THR HIS PHE PRO Peptide of Acetylcholine Receptor PHE
ASP GLU GLN Binds to B Cell MHC Class II. ASN CYS SER MET Activates
Helper T Cells, Stimulates LYS Autoantibody Production and
electrophysiologic signs of myasthenia gravis" by H. Yoshikawa, E.
H. Lambert, D. R. Walser-Kuntz, Y. Yasukawa, D. J. Mc Cormick and
V. A. Lennon in J. Immunol., 159 pp. 1570-1577 (1997)) j.
Acetylcholine receptor 67-75 (EMG) TRP ASN PRO ASP (see an article
entitled "The Main Region ASP TYR GLY GLY of the Nicotinic
Acetylcholine Receptor" VAL LYS by M. Bellone, F. Tang, R. Milius
and B. M. Conti-Troncoi in J. Immunol., 143 pp. 3568-3579 (1989))
k. Acetylcholine receptor 97-116 (EMG) ASP GLY ASP PHE (see an
article entitled "The Main Region ALA ILE VAL LYS of the Nicotinic
Acetylcholine Receptor" PHE THR LYS VAL by M. Bellone, F. Tang, R.
Milius and LEU LEU ASP TYR B. M. Conti-Troncoi in J. Immunol., THR
GLY HIS ILE 143 pp. 3568-3579 (1989)) 1. Acetylcholine receptor
195-212 (EMG) ASP THR PRO TYR (see an article entitled "Effect of T
cell LEU ASP ILE THR recognition and immunogenicity of TYR HIS PHE
VAL alanine-substituted peptides MET GLN ARG LEU corresponding to
97-116 sequence of the PRO LEU rat AChR alpha-subunit" by F. Baggi,
A. Annoni, F. Ubiali, R. Longhi, R. Mantegazza, F. Cornelio and C.
Antozzi in Ann. N. Y. Acad. Sci., 998 pp. 395-8 (2003)) (EMG) m.
Acetylcholine receptor 259-271 (EMG) VAL ILE VAL GLU (see an
article entitled "Peptide Analogs LEU ILE PRO SER to Pathogenic
Epitopes of the Human THR SER SER ALA Acetylcholine Receptor Alpha
VAL Subunit as Potential Modulators of Myasthenia Gravis" by E.
Zisman, Y. Katz-Levy, M. Dayanm, S. L. Kirshner et al in Proc. Nat.
Acad. Sci., USA, 93 pp. 4492-4496 (1996)) n. Acetylcholine receptor
129-145 (EMG) GLU ILE ILE VAL (see an article entitled "A 17-Mer
Self- THR HIS PHE PRO Peptide of Acetylcholine Receptor Binds PHE
ASP GLU GLN to B Cell MHC Class II. Activates Helper ASN CYS SER
MET T Cells, Stimulates Autoantibody LYS Production and
electrophysiologic signs of myasthenia gravis" by H. Yoshikawa, E.
H. Lambert, D. R. Walser-Kuntz, Y. Yasukawa, D. J. Mc Cormick and
V. A. Lennon in J. Immunol., 159 pp. 1570-1577 (1997)) o.
Acetylcholine receptor 67-75 TRP ASN PRO ASP (see an article
entitled "The Main Region ASP TYR GLY GLY of the Nicotinic
Acetylcholine Receptor" VAL LYS by M. Bellone, F. Tang, R. Milius
and B. M. Conti-Troncoi in J. Immunol., 143 pp. 3568-3579 (1989))
p. Sm B/B' protein (ELSE) (see an article PRO PRO PRO GLY entitled
"Side-Chain Specificities and MET ARG PRO PRO Molecular Modeling of
Peptide Determinants for Two Anti-Sm B/B' Autoantibodies" by J. A.
James, M. T. McClain, G. Koelsch, D. G. Williams, and J. B. Harley
in J. Autoimmunity, 12 pp. 43-49 (1999))
[0153] where: underlined amino acids make the most important
contributions to autoimmuneactivity; the table does not list all
the peptides which have been attributed to autoimmune disease
induction, however, this is a representative group; underlined
portions of some of the sequences refer to amino acids which are
critical for activity; and approximately 25-30 different
encephalitogenic regions have been described involving four (4)
central nervous system proteins--the ones listed are the most
carefully studied regions.
* * * * *
References